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Articles by C Gao
Total Records ( 4 ) for C Gao
  X Li , C Dong , S Shi , G Wang , Y Li , X Wang , Q Shi , C Tian , R Zhou , C Gao and X. Dong
 

Prion protein (PrP) is considered to associate with microtubule and its major component, tubulin. In the present study, octarepeat region of PrP (PrP51–91) was expressed in prokaryotic-expressing system. Using GST pull-down assay and co-immunoprecipitation, the molecular interaction between PrP51–91 and tubulin was observed. Our data also demonstrated that PrP51–91 could efficiently stimulate microtubule assembly in vitro, indicating a potential effect of PrP on microtubule dynamics. Moreover, PrP51–91 was confirmed to be able to antagonize Cu2+-induced microtubule-disrupting activity in vivo, partially protecting against Cu2+ intoxication to culture cells and stabilize cellular microtubule structure. The association of the octarepeat region of PrP with tubulin may further provide insight into the biological function of PrP in the neurons.

  M Guo , H Feng , J Zhang , W Wang , Y Wang , Y Li , C Gao , H Chen , Y Feng and Z. G. He
 

Sequence-specific DNA-binding transcription factors have widespread biological significance in the regulation of gene expression. However, in lower prokaryotes and eukaryotic metazoans, it is usually difficult to find transcription regulatory factors that recognize specific target promoters. To address this, we have developed in this study a new bacterial one-hybrid reporter vector system that provides a convenient and rapid strategy to determine the specific interaction between target DNA sequences and their transcription factors. Using this system, we have successfully determined the DNA-binding specificity of the transcription regulator Rv3133c to a previously reported promoter region of the gene Rv2031 in Mycobacterium tuberculosis. In addition, we have tested more than 20 promoter regions of M. tuberculosis genes using this approach to determine if they interact with ~150 putative regulatory proteins. A variety of transcription factors are found to participate in the regulation of stress response and fatty acid metabolism, both of which comprise the core of in vivo-induced genes when M. tuberculosis invades macrophages. Interestingly, among the many new discovered potential transcription factors, the WhiB-like transcriptional factor WhiB3 was identified for the first time to bind with the promoter sequences of most in vivo-induced genes. Therefore, this study offers important data in the dissection of the transcription regulations in M. tuberculosis, and the strategy should be applicable in the study of DNA-binding factors in a wide range of biological organisms.

  H Wang , A Chattopadhyay , Z Li , B Daines , Y Li , C Gao , R Gibbs , K Zhang and R. Chen
 

One of the key advantages of using Drosophila melanogaster as a genetic model organism is the ability to conduct saturation mutagenesis screens to identify genes and pathways underlying a given phenotype. Despite the large number of genetic tools developed to facilitate downstream cloning of mutations obtained from such screens, the current procedure remains labor intensive, time consuming, and costly. To address this issue, we designed an efficient strategy for rapid identification of heterozygous mutations in the fly genome by combining rough genetic mapping, targeted DNA capture, and second generation sequencing technology. We first tested this method on heterozygous flies carrying either a previously characterized dac5 or sensE2 mutation. Targeted amplification of genomic regions near these two loci was used to enrich DNA for sequencing, and both point mutations were successfully identified. When this method was applied to uncharacterized twr mutant flies, the underlying mutation was identified as a single-base mutation in the gene Spase18-21. This targeted-genome-sequencing method reduces time and effort required for mutation cloning by up to 80% compared with the current approach and lowers the cost to <$1000 for each mutant. Introduction of this and other sequencing-based methods for mutation cloning will enable broader usage of forward genetics screens and have significant impacts in the field of model organisms such as Drosophila.

  C Gao and A. Y. Wang
 

Human small intestine accounts for 75% of the gastrointestinal (GI) length but for only 1–5% of GI tumors. The reason remains as yet unclearly understood. Our study was designed to examine whether increased apoptosis and expression of related genes/proteins, especially those of the Bcl-2 family, contribute to this difference. For this purpose, 77 samples from patients were examined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and immunohistochemistry, including 40 cases from normal small intestine (jejunum), 7 cases from jejunum and ileum adenocarcinomas, and 30 cases from normal colon. The results showed that a significantly higher level of enterocyte apoptosis was observed in normal small intestine compared with small intestinal adenocarcinomas and normal colon (median of apoptotic index, 15.2% vs 0.1% and 1.6%, p<0.01). A similar pattern was observed for Bax (expression-positive, 77.5% vs 28.6% and 53.3%, p<0.05) but not for Bcl-2 (42.5% vs 42.9% and 46.7%, p>0.05) or Bax/Bcl-2 ratio (percent of samples having a ratio ≥1, 45.0% vs 14.3% and 36.7%, p>0.05). In conclusion, increased apoptosis and expression of Bax, not Bcl-2 or the Bax/Bcl-2 ratio, may play some role in the relatively lower incidence of human small intestinal carcinomas. However, more studies are required for a better understanding of these changes. (J Histochem Cytochem 57:1139–1148, 2009)

 
 
 
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