湖南农业大学学报（自然科学版）2016，Vol.42Issue(4)：424-428,5.
红鳍东方鲀去乙酰化酶 SIRT3基因的克隆及原核表达
Cloning and prokaryotic expression of silencing deacetylase SIRT3 gene from Takifugu rubripes
摘要
Abstract
In order to further study the function of gene SIRT3 in Takifugu rubripes, the full-length open reading frame (ORF) of gene T. rubripes SIRT3 (trSIRT3) was amplified from total RNA in their liver tissue through reverse transcription-polymerase chain reaction (RT–PCR). The recombinant expression vector pET32a/SIRT3 was generated from the prokaryotic expression vector pET32a (+) derived from sub-cloned ORF, then, it was transformed into E.coli Rosetta (DE3). As the result of IPTG induction, SIRT3 was successfully expressed in E.coli Rosetta (DE3). Result of sequence analysis showed that the ORF of SIRT3 was 1 263 bp and encoded with 420 amino acids. The prokaryotic expression system of recombined vector pET–32a/SIRT3 was successfully constructed. From the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the recombinant protein had an approximate molecular weight of 66 000, which was consistent with the theoretical molecular weight. The fusion protein with tag His, was mainly detected in the supernatant.关键词
红鳍东方鲀/去乙酰化酶/SIRT3 基因/基因克隆/原核表达Key words
Takifugu rubripes/silencing deacetylase/SIRT3 gene/gene cloning/prokaryotic expression分类
生物科学引用本文复制引用
张伟,姬明丽,张军华,千智斌..红鳍东方鲀去乙酰化酶 SIRT3基因的克隆及原核表达[J].湖南农业大学学报（自然科学版）,2016,42(4):424-428,5.基金项目
河南省教育厅重点项目(14A310027)；新乡医学院高学历人才启动基金(505001) （14A310027）
