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Fujun Zhou BS: Degree, University Contact Info: |
Description of Research
Mentor(s): Albrecht von Arnim
Rotation Summary
Mentor: Jizhong Zhou - Fall 2003
Study on Microbial Functional Gene Diversity for Belowground Ecosystem Using Clone-library and Microarray Approaches.
The objective of this project is to construct a clone libraies of gene amoA and pmoA for soil samples of OCCAM (Old-Field Community Climate and Atmosphere Manipulation)sample sites. The sequences of newlly identified genes will be used to design additional probes for a Functional Gene Array. Then use the new microarray to observe the diversity of underground microbiol community diversity. From 4 sites, A total of 59 amoA and 10 pmoA clones, respectively, were obtained, 3 different sequences were detected. For both amoA and pmoA clone libraries, low diversity was observed. The results of microarray showed that the majority (96.25%) of genes did not display significant changes among sample sites.
Mentor: Mitch Klebig - Spring 2004
RNA-based Gene-driven mutation scanning of the Cyropreserved Mutant Mouse Bank (CMMB).
The purpose of this project is to generate and optimize protocols for RNA-based gene-driven mutation scanning of CMMB (Cryopreserved Mutant Mouse Bank) samples. CMMB is a large collection of tissues and sperm from 4000 G1 male mice that are heterozygous for ENU induced mutations in Oak Ridge National Laboratory, life science division. Screening and studying the ENU caused gene mutations in mice helps us to elucidate the functions of human genes and proteins. However, for large genes with multiple introns, the traditional genomic DNA based mutation screening is time-consuming. we generated and optimized an effective largescale RNA-based gene-driven mutation screening system. Also screend 92 CMMB mouse RNA samples for mutations in Dnm2 and Pak4 genes. 12 putative mutations were detected by TGCE.
Mentor: Albrecht von Arnim - Summer 2004
Arabidopsis thaliana Genes CRY1, HY5 and PHOT1 full length gene-tagging for BRET.
BRET(Bioluminescence Resonance Energy Transfer) is a newlly developed approach to study protein-protein interactions in vivo. Compared to other protein interaction detecting approaches, BRET has the advantages of in vivo, in real time, inreal organism and nondestructive. However in Arabidopsis thaliana, the expression of RLUC fusion proteins (with 35S promotor, Gene ORF and RLUC) is not sufficent. Therefore it is necessary to generat a new gene tagging strategy for BRET. We generated a whole length gene tagging strategy. The fusion genes we use the native promotor instead of 35s promotor, also we included introns and 3' UTRs in our fusion gene. The tags (RLUC or YFP) was inserted to a position before the stop codon. We successfully tagged both RLUC and YFP to Arabidopsis thaliana CRY1 and HY5 genes, then cloned the fussion genes into pENTR/SD/D-TOPO vector.
Mentor: Robert Hettich - Summer 2004
Evaluation of methodologies for the Top-Down analysis of Rhodopseudomonas palustris
The purpose of this project is to evaluate the sensitivity accuracy and dynamic range of HPLC-FT-ICR-MS in topdown analysis. Two FPLC fractions of Rhodopseudomonas palustris proteins were examined to determine protein mass raange and PTMs using C4 and C5 reverse phase columns. 64 Proteins range from 5066.214 Da to 22237.314 Da of fraction 10 were observed C4 reverse phase column. 91 proteins mass range from 5075.675 Da to 23178.762 Da of fraction 11 were observed using C5 reverse phase column. Large proteins were separated better by C4 column, and small proteins were separated better in C5 column. Compared measured mass and calculated mass of 16 ribosomal proteins, 12 of them are within mass accuracy 10 ppm. Other than Methionine truncations, 17 putative protein PTMs were observed.