番杏Actin基因片段的克隆及生物信息学分析

本实验为研究番杏(Tetragonia tetragonioides)功能基因表达模式提供内参基因,根据登录在NCBI上植物的Actin基因的保守区域设计简并性引物,通过RT-PCR克隆获得番杏的Actin基因片段,将该片段连接于载体pGEM-T后进行测序,并将该基因序列通过生物信息学软件进行分析。结果显示,克隆所得基因片段大小为598 bp,编码198个氨基酸;该序列与登录在NCBI上的其他植物的Actin基因的核苷酸序列的同源性最大可达86%以上,编码蛋白的氨基酸序列同源性在88%以上。结果表明,本研究克隆所得的基因序列为番杏Actin基因片段。该基因命名为TtActin1,在Gen Bank的登录号为MH33308。

Cloning and bioinformatics analysis of Actin gene fragment from Tetragonia tetragonioides

In order to provide reference gene for studying the expression pattern of functional genes in Tetragonia tetragonioides, the cDNA fragment of Actin from Tetragonia tetragonioides was amplified by RT-PCR, by adopting degenerated primers designed according to the conserved region of the Actin genes which were submitted to NCBI database. The cDNA fragment was inserted into the pGEM-T vector and sequenced. The cDNA sequence was analyzed by online bioinformatics software. Result showed that this Actin cDNA includes 598 base pairs, encoding a partial actin with 198 amino acids. Sequence alignments showed that it shares over 86% nucleotide sequence identity and 88%amino acid sequence identity with other plant actins. This study indicates that a new Actin cDNA fragment is isolated from Tetragonia tetragonioides. This cDNA is named TtActin1 and submitted to GenBank(accession number: MH33308).