中国实验动物学报2011，Vol.19Issue(6)：456-460,5.DOI:10.3969/j.issn.1005-4847.2011.06.002
小鼠Lin28基因的克隆及原核表达
Cloning and prokaryotic expression of mouse Lin28 gene
摘要
Abstract
Objective Lin-28 is a gene recently shown to be involved in the conversion of somatic cells to induced pluripotent stem cells. The aim of this study was to clone Lin28 gene and analyzed its expression in E. Coli so to make an effort on the further study of its function. Methods Lin28 gene was cloned from the cDNA amplified from 8. 5-day mouse embryos mRNA, and ligated with linear vector pMD18-T. The recombinant plasmid was digested and the segment with Lin28 gene was purified then subcloned into prokaryotic expressing vector pET-30a(+). Results The recombinant plasmid was transfected into E. Coli strain Rosetta(DE3) and its expression induced by IPTG was identified as well analyzed by SDS-PAGE. Compared with the DNA (NM-145833) , the homology was 99. 37% , and compared with the amino acid sequence, the homology was 100%. Conclusions The coding region of Lin.28 in a length of 630 bp is successfully harvested and recombinant protein with 27. 5 kD just as anticipated.关键词
ICR小鼠/Lin28/克隆/原核表达Key words
ICR mouse/ Lin28 gene/ Clone/ Prokaryotic expression分类
生物科学引用本文复制引用
宁方勇,王春生,张秋婷,安星兰,朴善花,安铁洙..小鼠Lin28基因的克隆及原核表达[J].中国实验动物学报,2011,19(6):456-460,5.基金项目
国家自然科学基金资助项目(30771538,31000990) （30771538,31000990）
中央高校基本科研业务费专项资金资助(DL10BA06). （DL10BA06）
