CLONING AND EXPRESSION OF THE FRAMESHIFT Zbtb7A PROTEIN FROM CARCINOGENESIS TISSUE IN E. coli
Zbtb7A, a transcriptional repress factor, plays a key role in cancer pathogenesis. It is overexpressed in multiple human cancers and cells lacking this factor are refractory to oncogenic transformation. We recently detected a frameshift mutation of Zbtb7A gene in tumor tissue from Chinese patient and tumor cell line. In the frameshift Zbtb7A gene, excluding for the POZ domain conserved, a nuclear localization signal at the C-terminus was predicted according to bioinformatics methods. To obtain the frameshift protein we tried to clone and express this mutant in the E. colt. The cDNA was obtained by RT-PCR from gastric carcinoma patient s carcinogenesis tissue and cloned into pMD 19-T Simple vector. After sequencing the target fragment was inserted into the E. coti expression vector pGEX4Tl and expressed in E. coli Rosetta (DE3). Induced with IPTG and detected with 10％ SDS-PAGE, a band corresponding to a protein of 62KD was detected. The protein was distributed mainly in the insoluble fraction. This gene as well as its protein may become a promising biomarker or target for tumor diagnosis and therapy.
Zbtb7A carcinogenesis tissue frameshift Rosetta (DE3) pGEX4T1
Hui Wang Qingling Wang Yahong Huang Shuli Zhao Yayi Hou
Immunology and Reproductive Biology Lab, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210093, China
国际会议
The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)
杭州
英文
91-94
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）