Human interleukin-24 cDNA cloning and prokaryotic expression of mutations to correct.docVIP

 PAGE \* MERGEFORMAT 12 Human interleukin-24 cDNA cloning and prokaryotic expression of mutations to correct Author: Weili Li, Cheng-Hua Li, Yuan Chengfu, Chen Ji, Song Ding Tao, Liu Ge Li, Yi Fa-Ping, SONG Fang-zhou [Abstract] Objective To obtain human interleukin 24 (IL  24) cDNA, construct prokaryotic expression vector to express glutathione S-transferase containing (GST) fusion protein GST  IL  24. Approach to human peripheral blood mononuclear cells (PBMC) total RNA as template, RT  PCR amplification of IL  24 cDNA, was cloned into the prokaryotic expression vector pGEX  4T  2, sequencing results showed that IL  24 cDNA No. 223 GA, 370 TC, thereby leading to protein-coding amino acid changes: A75T, H124Y, through secondary PCR be corrected, the recombinant plasmid pGEX  IL  24 transformed into E. coli BL21 (DE3), isopropyl  β  D  sulfur on behalf of the thiogalactoside (IPTG) induced expression, SDS  PAGE, Western blot test showed that the fusion protein GST  IL  24 expression. Results by RT  PCR and secondary PCR to be human IL  24 cDNA, to build its prokaryotic expression vector by IPTG induction in relative molecular weight of about 50 000 a fusion protein. Conclusion IL  24 of the recombinant vector in E. coli BL21 (DE3) to express GST  IL  24 fusion protein. [Keywords:] interleukin-24; second PCR; mutation corrected; fusion protein; cloning; prokaryotic expression Interleukin-24 (interleukin  24, IL  24) was in 1995 by Columbia University, Jiang et al [1] The study found, and it can selectively inhibit the growth of broad-spectrum cancer cells, promote apoptosis, and has a certain degree of immune regulation. The replication-deficient adenovirus carrying IL  24 has entered phase II clinical trial results showed: a significant effect of the very few toxic side effects [2]. Thus, IL  24 gene therapy is expected to become a new candidate genes. We intend by RT  PCR method to get IL  24 cDNA, to b