菘蓝APX基因克隆及序列分析

Cloning and identification of APX gene from Isatis indigotica

关键词:  菘蓝  APX基因  真核表达载体  基因克隆

Key words:  Isatis indigotica  APX gene  eukaryote expression vector  gene cloning

摘要:

为进一步研究菘蓝APX基因的功能，构建了菘蓝APX基因真核表达载体。从菘蓝植株中提取总RNA，逆转录为cDNA，根据APX基因在该类植物中的同源性设计简并引物，利用PCR方法钓取目的基因，将目的基因与T载体连接，PCR检测阳性克隆，同时菌液送往测序公司进行测序。结果表明：测序片段的生物信息学分析证实了该序列与GenBank登录的APX基因一致。说明成功构建了菘蓝APX重组质粒和克隆鉴定了菘蓝APX基因，可进一步用于基因表达和表达产物的功能研究。

Abstract:

To construct the recombinant eukaryotic expression plasmid of pTZ57R/T/APX. APX gene was amplified from the genomic DNA of Isatis indigotica by polymerase chain reaction(PCR). APX gene was then inserted into pTZ57R/T eukaryote expression vector. The positive colonies were screened and identified by PCR 〖JP3〗and sequencing. The specific gene APX which was about 1 257 bp，was successfully amplified and pTZ57R/T APX were constructed. The DNA sequence of APX gene was as the same as the APX gene nucleotide sequences published in Genebank. pTZ57R/T APX recombinant was successfully constructed. And the construction provides the basis for the further study.