安徽农业科学2012，Vol.40Issue(3)：1324-1328,1331,6.
蜡梅花香基因SAMT的cDNA克隆及在大肠杆菌中的表达
cDNA Cloning and Expression of SAMT Gene from Chimonanthus praecox in Escherichia coli
马蕾 1李慧芬 1彭忱晨 1陈子柱 1龙章富1
作者信息
- 1. 四川大学生命科学学院生物资源与生态环境教育部重点实验室,四川成都610064
- 折叠
摘要
Abstract
[Objective] The aim was to study the cDNA cloning and expression of SMAT gene from Chimonanthus praecox. [Method] A novel cDNA was cloned by PT-PCR using the total RNA as template,and amplified to the desirable size. The RT-PCR products were reclaimed and transformed into E. Coli DH 5a together with the PMD18-T vector after ligating by T-A cloning. Identified by colony PCR and EcoRI and NotI digestion,the recombinant plasmid with target gene was screened out and conducted the sequence analysis. [Result] Results of the sequence analysis showed that the ORF fragment of the SAMT cDNA was successfully cloned form Chimonanthus praecox gene,with the length of 1 196 bp and encoded 380 amino acids fragment which shared 99. 2% homology to that of previously reported SAMT cDNA from Chimonanlhus praecox (ABU88887). The SAMT cDNA fragment was sub-cloned into the prokaryotic expression vector PGEX1-4T-1 ,and the obtained recombinant plasmid was named PGSAMT. After inducting by 0. 0lmol/L IPIG,the result of the SDS-PAGE analysis showed that the molecular weight of the fusion expression SAMT protein was about 66 kDa, which was close to the predicted fusion protein derived from the 26 kDa GST band and 42. 3 kDa SAMT gene of Chimonanthus praecox encoded protein. [Conclusion] This study successfully cloned and expressed the SAMT gene of Chimonanthus praecox.关键词
蜡梅/花香基因/cDNA/克隆/原核表达Key words
Chimonanthus praecox/ SAMT/ cDNA/ cloning, prokaryotic expression分类
生物科学引用本文复制引用
马蕾,李慧芬,彭忱晨,陈子柱,龙章富..蜡梅花香基因SAMT的cDNA克隆及在大肠杆菌中的表达[J].安徽农业科学,2012,40(3):1324-1328,1331,6.
