STEM Teacher and Researcher (STAR) Program Posters

Molecular Modeling of Novel Fuel Cell Membranes

Tessa Brown et al. — Mon, 28 Jan 2013 13:59:14 PST

Fuel cells are clean and efficient energy conversion devices. The fuel cells of interest contain polymer electrolyte membranes (PEMs) that inhibit the conduction of electrons and facilitate the transport of protons. Nafion® is the most widely used membrane for fuel cell applications. However, alternatives are desired because Nafion is expensive, allows significant amounts of methanol crossover, and functions poorly at low humidity or high temperature. An acid-base blend membrane composed of both acidic sulfonated poly(ether ether ketone ketone) (Ph-SPEEKK) and basic polysulfone tethered with 5-amino-benzotriazole (PSf-BTraz) has been show to perform better than traditional acidic PEMs such as Nafion and Ph-SPEEKK. We use molecular dynamics to study the PEM morphology and the transport of water, hydronium, and methanol in Ph-SPEEKK/PSf-BTraz blend membranes. Our aim is to understand the fundamental science behind the enhanced properties of the blend membrane. Initial analysis shows that transport is slower and sulfonate groups are farther apart compared to plain Ph-SPEEKK. The decrease in methanol crossover may account for the enhanced performance Ph-SPEEKK/Psf-BTraz. Further analysis is needed to definitively relate the structure to transport properties.

Steroids and SSRIs in Wastewater and EE2 Uptake in FHM

Elizabeth Ellen Wittenberg et al. — Thu, 24 Jan 2013 15:49:06 PST

Pharmaceuticals have been found in waste water treatment plant (WWTP) effluents and there are scientific studies that indicate at least some of these chemicals have adverse effects on fish health. Two groups of pharmaceuticals that currently cause concerns are natural and synthetic steroids and selective serotonin reuptake inhibitors (SSRIs). Fathead minnows (FHM) are a common species of fish used for research and ethynyl estradiol (EE2) is a synthetic estrogen known to cause decreased fertility in fish. FHMs were exposed to EE2 for time periods between 0.25 hours to 144 hours in order to study uptake and disposition in this species. These results demonstrated that EE2 is rapidly absorbed by FHMs and distributed to target tissues such as the liver and gonads. These findings support the use of short-term exposure studies of WWTP effluents to assess the potential physiological effects on fish. The use of relatively short exposure times (e.g. 72 hrs) requires fewer resources and permits more WWTP effluents to be tested.

Further Development in Nondestructive Methods to Gauge Life Expectancy in Ferromagnetic Components

David Millard et al. — Thu, 24 Jan 2013 13:54:22 PST

Manufactured nuclear components under stresses induced through normal operations cause mechanical fatigue and strain. Depending on their magnitude and distribution they can contribute to increasing the expected life of a component or for its premature failure. Using Barkhausen noise we can analyze the microstructural characteristics without damaging the sample through magnetization or acoustics. The samples in our case are ferromagnetic metals, also known as ferrous metals, from heat treated and rolled steel. A Rollscan 300 instrument and Microscan 600 software were used to acquire Barkhausen noise data from fatigued steel samples. MATLAB software and R software were used to evaluate results of the Microscan 600 to better understand the signal processing algorithms. In order to find a correlation we used a two random variable probability distribution function (PDF). plot We found the difference between the three positions taken on the given sample at each strain level, and with a 95% confidence level we created a plot of data points that found a loose correlation in the data results between both perpendicular and parallel testing. Using these results we can compare older sets of data and create an accurate prediction of stress levels induced upon nuclear components. We hope to create more precise predictions in the near future using alternative methods, such as statistical calibration techniques to find closer one‐to‐one correlations.

Characterization of Coke from the Catalytic Pyrolysis of Biomass

Kristen Ryan et al. — Wed, 23 Jan 2013 13:34:02 PST

The conversion of non‐food sources of biomass to liquid transportation fuels could be a viable alternative to non‐renewable petroleum based fuels with high carbon dioxide emissions if the process can become cost effective. One of the problems with the catalytic pyrolysis as a thermochemical pathway to biomass based fuel production is the accumulation of carbonaceous deposits (aka coke) that can completely deactivate catalysts over time. A multiple technique approach was used to analyze four different stages of coke development. Solid samples of coke were analyzed via multiple techniques; elemental, 13C NMR, TGA, and SEM analysis confirmed the presence of two types of coke molecules and low (.5) hydrogen to carbon ration at all stages of deactivation. This suggests that coke is very similar to coal and could potentially be intercepted by hydrogen donor molecules in the pyrolysis process to produce more desirable hydrocarbon molecules. GC/MS results from carbon‐tetrachloride (CCl4) and methylene‐chloride (CH2Cl2) extractions indicate that coke molecules formed during fast pyrolysis (500‐600°C) are insoluble which was unexpected.

Using Modeling and Simulation to Analyze Complex Aircraft

Kimberlee Margosian et al. — Wed, 23 Jan 2013 12:52:12 PST

Modeling and Simulation (M&S) is used at the Air Force Flight Test Center (AFFTC) on Edwards Air Force Base (AFB) to better understand physical phenomena on aircraft. M&S allows for the reduction of cost and risk by providing a better understanding of required flight tests and the interactions between various forces and the aircraft (i.e. wind resistance, pressure change, and temperature change). Without this process, the lives of pilots would be at a much greater risk when testing their aircraft and there would be little to no funds to fly due to the cost to repair or modify the aircraft. This process uses aero-structure interaction simulation and fluid mechanics simulation data measuring wing tip flutter to characterize and correlate the simulated findings on complex aircraft configurations. The data that has been gathered can be analyzed with Microsoft Office Excel’s data analysis function. Fourier analysis is performed on the Z coordinate of each data set to help measure amplitude versus frequency based off data measuring deflection versus time. The findings have shown the peak frequency varies based on the time interval and amount of data points tested. With a smaller time interval and more data per period, there is more accuracy in determining the shape of the graph but variance in peak frequencies. With a larger time interval and less data per period, determining the true peak frequency is more effective.

Chloroform Extraction of Iodine in Seawater Method Development

Hanna Seidler et al. — Thu, 06 Dec 2012 14:24:22 PST

The extraction of iodine from seawater is used as a means of analyzing the concentration and isotopic ratios of iodine at different locations in the ocean. This has practical applications in the testing of discharge from nuclear fuel reprocessing plants, tracing of ocean currents, and testing areas for potential environmental and health impacts. One of the current methods used is separation extraction involving chloroform (CHCl3). This method is lengthy (almost an hour per sample) and does not guarantee 100% recovery of the iodine in the water. This research seeks to optimize the existing protocol for efficiency while maintaining or improving recovery. We assessed each methodological change qualitatively using a color scale (I2 in CHCl3) and quantitatively using Inductively Coupled Plasma Mass Spectrometry (ICP‐MS).

The effects of low dose gamma radiation on the production of bone cell communication factors

Anna DeNeui et al. — Mon, 03 Dec 2012 14:34:04 PST

The health of skeletal tissue is dependent, in part, on a balanced coexistence between two main cell types: osteoblasts, which form the mineralized structure of bone, and osteoclasts, which digest and break down bone. Communication between these two cell types is facilitated by osteoblast production of receptor activator of nuclear factor kappa‐B (RANK) ligand and osteoprotegerin (OPG). The increased bone loss anticipated after long‐term spaceflight may be associated with the degenerative effects of ionizing radiation on this system. We hypothesize that ionizing radiation associated with spaceflight modulates RANK‐Ligand/OPG expression in osteoblasts, consequently altering the behavior of osteoclasts. To examine the effects of irradiation on mRNA levels of RANK‐Ligand/OPG, and subsequent effects on osteoclasts, we irradiate MC3T3‐E1 cells (osteoblast cell line) with 2Gy of ionizing radiation and introduce RAW 246.7 cells (osteoclast cell line) to the culture 1 day after. Q‐PCR is performed for RANK‐Ligand and OPG gene expression at multiple time points to assess for changes in RANK‐Ligand/OPG ratio. Calcium phosphate resorption plates as well as TRAP staining are used to assess osteoclast levels of differentiation and bone resorption. These results increase our understanding of the implications of ionizing irradiation for human bone health, further preparing us for long‐term spaceflight.

To Mask or Not to Mask: Using a Model‐Based Barameter to Detect Vegetation Changes from Polarimetric and Interferometric Radar Data

Kaitlyn Fiechtner et al. — Mon, 03 Dec 2012 14:24:55 PST

This project focuses on implementing a software component that will help the NASA JPL (Jet Propulsion Laboratory) UAVSAR (Uninhabited Ariel Vehicle Synthetic Aperture Radar) group ultimately estimate Earth’s biomass and vegetation properties. Estimating the worldwide biomass will contribute to our understanding of the changing climate. By utilizing Cloude’s model-based incoherent decomposition we are able to extract the maximum surface-to-volume scattering ratio from the polarimetric and interferometric SAR (Pol-InSAR) data. The surface-to-volume ratio contains information about the structure of the imaged vegetated area, and can be used to detect vegetation changes between data acquisitions. The software component will create an image mask using a threshold based on the surface-to-volume ratio. The mask will be needed at times when vegetation changes between data acquisitions are too large to allow for accurate estimation of forest parameters from Pol-InSAR data. In order to implement the mask, we implemented the mathematical model in C programming language and incorporated it into the parameter estimation program. In the presentation we will describe the theoretical model and show results of the estimation surface-to-volume ratio on using UAVSAR collected over the Harvard Forest (MA).

Building the Foundation for Student Observations Using NASA's Deep Impact Spacecraft

Jamie Vargas et al. — Mon, 03 Dec 2012 14:24:38 PST

On July 4, 2005, NASA’s Deep Impact spacecraft successfully released an impactor into the nucleus of comet Tempel 1 in hopes of answering the fundamental question of what makes up the composition of a comet. Upon the completion of its primary mission, NASA approved a second mission, EPOXI, with a dual purpose to study extra‐solar planets as well as comet Hartley 2 using the still‐flying Deep Impact spacecraft . Today, a team of scientists and engineers are again proposing to perform a third mission – Deep Impact 3 (DI3) using the Deep Impact spacecraft. As part of the Education and Public Outreach (E/PO) initiative of the mission, the team has designed a community science program designed primarily for high school and higher education students to conduct real life – real time experiments and research. My contribution to this proposed mission consists of three components. The first component is designing a tutorial for acquiring and processing data from the DI Science Team’s archive so that students and educators can process and calibrate raw data into a final processed image. The second component is a lesson plan that uses several light curves produced from EPOCh (Extra‐solar Planet Observations And Characterization), to explore Kepler’s Laws of Orbiting bodies. The final component of my project is to design a science campaign that will take images of an astronomical object that is within the boundary conditions of the spacecraft. The purpose of the scientific campaign is to allow new data to be accessible to the general population as well as provide an opportunity to share the data with my physics students upon my return to the classroom.

Impact of Neutron Irradiation on the Thermoelectric Properties of Rare Earth‐Based Thermoelectric Materials

Zachary Chandler et al. — Mon, 03 Dec 2012 14:24:34 PST

The impact of neutron irradiation on the thermoelectric (TE) properties of n‐type La 3‐x Te 4 , p‐type Yb 14 MnSb 11 , and n‐ and p‐type filled skutterudites is reported and discussed. During operation in Radioisotope Thermoelectric Generators (RTGs), these TE materials are expected to experience a certain degree of radiation over time, specifically, fission neutrons. This could affect their TE properties and thus the performance of the generators over time. In this study three samples of each of the above materials were exposed to 18 years worth of neutron radiation near room temperature at the Ohio State University Research Reactor (OSURR). Their electrical resistivity, carrier mobility, carrier concentration, thermal conductivity, Hall coefficient, and Seebeck coefficient were measured before and after radiation exposure at room temperature. Post‐irradiation the properties are measured twice. The first measurements were conducted on the samples as received after irradiation. The samples were then polished to remove any surface discoloration or oxidation films. The above properties were tested again to determine if the initial surface characteristics played an influence on the TE properties. The room temperature TE properties indicate that the neutron exposure had limited impact on the Seebeck and resistivity (less than 10% deviation). Results agree with previous investigations that established the minimal impact of neutron exposure on other similar TE materials. High‐temperature TE property measurements, including thermal conductivity, will also be performed on the samples to confirm the initial room temperature results.

Electrical Energy Storage: Synthesis of Metal Oxide Nanostructures for Potential Use in Flow Cell Batteries

Robin Young et al. — Mon, 03 Dec 2012 14:24:32 PST

One of the major concerns for the near future is our ability to generate and store energy. Though green technologies for energy generation such as solar and wind harvesting have been developed, their constant fluctuation in output requires a cost‐effective and efficient mode for storing the energy that is generated. The flow cell battery was developed by NASA in the 1970’s and has recently garnered attention in Japan, Australia, and California for large scale energy storage. Flow Cell batteries were originally produced using Iron and Chromium compound electrolytes; however the relative abundance, multiple valence states, and low toxicity of Vanadium make it an excellent alternative candidate for an electrolyte. The current Vanadium Redox Batteries use Vanadium ions dissolved in sulfuric or hydrochloric acid, and one drawback is that the particles tend to aggregate in the filters of the batteries, which decreases lifetime. I plan to synthesize Vanadium nanostructures, such as nanowires, for use in these batteries with the hopes of increasing mass and electrical transport efficiency and preventing the buildup of particles within the devices.

Propidium Monoazide qPCR for Viability Assessment of Microbes with Species Specificity

Carolyn Laymon et al. — Mon, 03 Dec 2012 14:24:30 PST

Currently, there are no portable instruments capable of both pathogen viability assessment and microbial identification. There is an urgent need to distinguish between viable and inactivated microorganisms, as well as assess if they are pathogenic. Propidium monoazide (PMA) is a DNA intercalating dye that is cell membraneimpermeable, thus it only comes in contact with the DNA of dead cells. Polymerase chain reaction (PCR) amplifies specific sequences of DNA allowing for the precise identification of organisms, such as pathogens. Through binding of DNA, PMA strongly inhibits PCR amplification of dead cells, which enables the selective DNA‐based detection of viable pathogens. Therefore, a PMA‐PCR combined assay, rather than conventional PCR, has the potential to distinguish between specific viable and nonviable bacteria or viruses. However promising, there is a lack of evidence demonstrating PMA’s use as a true quantitative tool for viability assessment, which will be critical for most real‐world applications. This stems in part from: i) the lack of a rigorous comparison of PMA‐qPCR with gold standards such as culturing, and ii) a lack of information on how PMA enters cells and inhibits PCR. To elucidate the quantitative effectiveness of PMA‐qPCR, mixed fractions of viable (non‐heat –inactivated) and dead (heat‐inactivated) E. coli K12 strain cells were prepared (Viable:Dead, 100:0, 75:25, 50:50, 25:75, 10:90, 1:99, 0:100) and examined through PMA‐qPCR, as well as fluorescence microscopy and cell culturing for comparison against conventional methods. We also assessed the mechanism of PMA permeability through PMA‐qPCR of E. coli inactivated by mechanisms independent of membrane permeation. We found that PMA‐qPCR is only quantitative at 10% and lower viable proportion of cells in a sample, and that the PMA mechanism of action may be part precipitation, as well as an inhibitor of polymerase processivity.

Raman Mapping of Mars Relevant Minerals Using Multiple Excitation Wavelenghts

Victoria Campbell et al. — Mon, 03 Dec 2012 14:24:25 PST

The purpose of this project is to map Mars relevant minerals using the Raman instrument Horiba LabRam Raman. The Raman spectrometer is a non-invasive sampling technique used to identify mineral composition of various compositions by measuring the “Raman Shift” of each mineral. This was accomplished by using a green (532nm) laser which acted as a monochromatic light source. This source was absorbed by the sample and reemitted. The frequency of the reemitted photons are then shifted up or down in comparison with original frequency, which is referred to as the ‘Raman Effect’. The samples with shorter wavelengths produced a stronger Raman effect with easily identifiable excitation wavelength peaks. The current project focuses on refining this technique to identify minerals by examining minerals of different compositions using various magnification objectives in conjunction with various filters to simulate variation in spot size and power density put down on the sample. The results of this project have yet to be analyzed and the data for the red (632nm) laser and a UV laser (325 nm) still needs to be collected. By the end of this study, we expect to learn which monochromatic light source works best at identifying minerals of different compositions. This work is foundational to the development of the fiberless-Raman Fluorescence micro-mapping instrument.

Screening the Farallon Island Bacterial Collection for Ionic Liquid Tolerance to Create Genomic Libraries

Alison Richins et al. — Mon, 03 Dec 2012 14:24:18 PST

To find a gene that confers ionic liquid resistance in bacteria, environmental samples of bacteria were taken from a high salt environment in the FarallonIslands. The bacteria were grown on different media that contained either 1‐Ethyl‐3‐methylimidazolium chloride (EMiM‐Cl) or 1‐Ethyl‐3‐ methylimidazolium acetate (EMiM‐Ac) to determine which bacteria should be used to create a genomic library to screen for genes conferring ionic liquid resistance. Bacterial isolates IL 210, IL 15, CX32 AND CX 38 grow on media containing EMiM‐Cl as well as media containing EMiM‐Ac and are recommended for the creation of a gene library.

Evaluation and Calibration of the Focusing, and Tilting and Chopping Mechanisms of SOFIA

Brian Eney et al. — Mon, 03 Dec 2012 14:24:15 PST

The Stratospheric Observatory for Infrared Astronomy (SOFIA) is a 2.7 meter telescope on board a Boeing 747‐ SP. The telescope has been receiving upgrades to equipment and instruments, as a result the Telescope Assembly needs to be evaluated and calibrated to achieve optimal image quality. The challenge to collimating a 2.7 meter telescope without special equipment attached to the telescope is essential for optimal observation time. This calibration needs to done in flight with science instrument in place and take only minutes to perform for maximum observing time and data collecting.

Unmasking the Mysteries of High-Mass X-Ray Binaries (HMXBs): The Role of LLNL’s Electron Beam Ion Trap (EBIT)

Carey Baxter et al. — Wed, 14 Nov 2012 15:18:54 PST

The Electron Beam Ion trap (EBIT) uses a very narrow electron beam (~60µm) to excite and trap ions. X-ray emissions of the excited ions are then diffracted and analyzed. I studied specific spectral emission lines of ionized silicon. This data can be used as a point of reference for similar spectra measured by the satellite Chandra so that the Doppler shift due to wind around the accretion disks of High Mass X-ray binaries (HMXBs) can be calculated. HMXBs are pairs of stars that are luminous in X-rays. They are composed of a donor star that gives up mass to an accretor, a compactor but more massive star like a neutron star or black hole. Knowing more about the wind movement around HMXBs helps us to better understand the lack of uniformity of mass observed in accretion disks.

Survival of B. horneckiae Spores Under Ground-Simulated

Sneha Tharayil et al. — Wed, 14 Nov 2012 10:36:19 PST

Increasing efforts to find and study habitable conditions and the possibility of life on extraterrestrial cosmic bodies have also deepened the need for planetary protection advancements to prevent possibilities of forward and backward contamination. Despite stringent precautions to ensure spacecraft cleanliness, a number of spore-forming bacteria, including the biofilm forming strain, Bacillus horneckiae have been isolated from spacecraft associated surfaces. B. horneckiae spores have demonstrated unusually high resistance to rigorous sterilization procedures, and extreme conditions, including intense UV radiation. This exceptionally high tolerance of extreme conditions demonstrated by spore-forming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. Thus, similar to prior EXPOSE projects commissioned by ESA, the BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, are potentially able to withstand harsh space conditions.

The objective of this study was to establish reference data on the survivability rates of B. horneckiae spores after exposure to certain ground-simulated space conditions: UV-space radiation, dark-space (no) UV radiation, simulated Martian UV, and simulated dark-Martian UV radiation. B. horneckiae spores were extracted from metal coupons using polyvinyl alcohol, after which serial dilutions were performed to determine the total number of viable spores in each coupon. The results showed that simulated UV-space exposed spores had a lower survivability rate than simulated dark-space exposed spores. Similarly, simulated Martian UV had a greater reduction effect on spore survivability than simulated dark Martian UV conditions. These results potentially hold strong implications for predicting the survivability of B. horneckiae communities in real space.

Playing "Hide and Seek" With Binary Stars

Elin Deeb et al. — Wed, 14 Nov 2012 10:36:12 PST

Asymptotic Giant Branch (AGB) stars are in a later stage of life than stars like our Sun. these stars are beginning to shed their outer layer and will eventually form planetary nebulae (PNe). The evolution of a PN impacts the evolution of the surrounding galactic environment, but PNe often have asymmetric geometries which are not entirely understood. The influence of a binary companion is predicted to affect the evolution of a PN although it is difficult to detect companions around AGB stars due to their dusty envelopes, large luminosities and variability. material shed by AGB stars however would likely form a disk of matter surrounding the companion, much as is observed in the mira system. if the companion is hotter than the GB star, the radiation from the companion would cause the disk to emit far ultraviolet (FUV) radiation in excess of what would be given off by an AGB star (and its surrounding envelope) due to the AGB star's lower temperature. 12 objects were identified for study based on UV, IR and optical blackbody data which indicated the likely presence of hotter companion stars. Spectroscopic UV data was then obtained by GALEX to search for characteristic spectral features typical of known AGB binary systems. The resulting "peek" into these systems however is relatively inconclusive due to missing FUV data for all but one object (resulting from an instrument failure) combined with the lower resolution of the GALEX grism. Future observations by the Cosmic origins Spectrograph (COS) on the Hubble Space Telescope are recommended to further prove these systems.

Further Development of the Chemical Mixture Methodology (CMM) Wizard

Sarah Horn et al. — Mon, 05 Nov 2012 11:43:11 PST

Title: Further development of the Chemical Mixture Methodology (CMM) Wizard – an on-line tool

Sarah Horn, University of Colorado, Boulder

The Chemical Mixture Methodology (CMM) program was developed to ensure individuals’ safety in the event of a chemical release into the atmosphere. The CMM uses Health Code Numbers (HCN) and user entered chemical concentrations to calculate hazard indices (HIs). The HIs indicate the level of adverse health effects an individual could experience from exposure to a mixture of chemicals. Current focus is on furthering development of the CMM Wizard, an online application that allows a user to input information about a specific chemical mixture (i.e. chemical names, receptor locations, concentration values) and then calculates the HI for the mixture. The Wizard then displays the results based on the mode of action of the chemicals and the target organs effected by the chemicals. In order to further develop this program, multiple tests have been executed to determine the Wizard’s accuracy and benefit as evidenced by the level of max HI as compared to simply adding HIs for each chemical. The Wizard test results were compared to hand calculations to ensure accuracy. Testing of CMM Wizard is still in progress. Apart from assisting with analysis of the CMM Wizard, other project goals were to compose a user guide and develop a contact page using HTML for the CMM Wizard. One additional goal was to create a user registration page for the CMM using HTML. The CMM Wizard user manual provides users with an easy to understand workable instruction guide along with troubleshooting tips. The CMM Wizard contact page allows users to report problems, pose questions, or submit comments. The registration page allows CMM users to register and receive updates on CMM tools and progress. In the future, a beta version of the CMM Wizard will be released with the user manual and new features.

Improving the Efficiency of Homologous Gene Replacement by Disrupting Non-Homologous End Joining Pathway Gene KusA in the Oleaginous Fungus Mortierella alpina

Kathleen T. Krueger et al. — Fri, 02 Nov 2012 10:53:55 PDT

Mortierella alpina , a oleaginous filamentous fungus, is one of industrial fungal strains known for the production of arachidonic acid. It is also of particular interest for hydrocarbon biofuel production since it is able to produce up to 50% of its mass in rich, long-chain polyunsaturated fatty acids [PUFA’s]. In addition to high fatty acid production, M. alpina like many other oleaginous fungi, already have mechanisms for accumulating significant concentrations of hydrophobic compounds making it a naturally equipped candidate to handle potential toxic concentrations of hydrocarbons. To date, three different transformation methods have been established for M. alpina that are capable of sporulation. These are particle bombardment, chemical, and Agrobacterium-mediated transformations. However, many M. alpina strains, such as M. alpina ATCC3222, are poor or absent of sporulation, of which have only protective coats, and the much less penetrable plant cell wall, yet remain of interest for their high lipid production. The goal of this study was to develop an efficient transformation method for those strains, hence allowing researchers to further manipulate these fungi for further improvement of lipid production. Included was optimization of best culture medium for growth and maintenance, optimal conditions for protoplast generation, and replacement of the homologous KusA gene. In molecular principle, homologous recombination is the most efficient method of disrupting, modifying, or replacing a target gene. This integration of exogenous DNA does not come readily to most organisms. Capitalizing from prior research efforts, it is known that deletion of KusA increases the rate of homologous recombination up to 100% in Neurospora strains (Ninomiya 2004).The kusA gene deletion vector, comprised of three different DNA fragments, namely the upstream region of KusA gene, the antibiotic selection marker and downstream region of KusA gene, was assembled by yeast-gap repair method. The gene deletion vector yeast genomic DNA mixtures were rescued by E. coli transformation. The isolated plasmid DNA was confirmed by PCR and digested with endonuclease restriction enzyme PmeI for M. alpina transformation. The protoplast isolation was optimized by adjustment of the enzymes concentration and protoplast buffers. The optimization of protoplast transformation is still undergoing. A successful deletion of KusA gene within biotechnologically important M. alpina could enable homologous recombination of other genes of interest in a higher frequency. This capacity may also improve the advancing the production of microbial oils for bioenergy and arachidonic acid human health applications.