芸芥果胶酸裂解酶基因的克隆及序列分析

采用RACE技术从芸芥自交亲和系花药中克隆出果胶酸裂解酶(Pectate lyases,PL)基因,全长1 657bp,其完整开放阅读框(Open Read Frame,ORF)为1 371bp,编码456个氨基酸,分子质量51.179ku,等电点9.42。序列比对结果表明,该蛋白与其他植物的PL蛋白具有较高的同源性,因此命名为EsPL。进化树分析表明,芸芥EsPL基因与十字花科芸薹属植物甘蓝型油菜、白菜型油菜的PL基因亲缘关系较近。实时荧光定量PCR结果显示,EsPL在芸芥开花后自交亲和系花药中的表达量显著高于自交不亲和系花药中表达量,该基因可能在芸芥自交亲和性状调控过程中发挥重要作用。

Cloning,Sequence Analysis of EsPL Gene from Eruca sative Mill

The full-length cDNA of EsPL was obtained by the technology of rapid amplification of cDNA ends (RACE).The full-length cDNA was 1 657 bp, containing a 1 371 bp opening reading frame(ORF).The deduced protein was 456 amino acids with molecular mass 51.179 ku and isoelectric point 9.42.The sequence aligment demonstrated that EsPL was high identity with other PL proteins from other species.So it was named the EsPL.Phylogram tree showes that there were closest evolutionary relationship of Brassica napus, Brassica rapa and Eruca sative Mill gene.Analysis by real-time PCR indicated that EsPL gene expression levels in SC anther is significant higher than SI anther after flowering,which suggested that EsPL played an important role in SI and SC characteristics of regulation of Eruca sative Mill.