大青杨纤维素合酶PuCesA7基因片段的克隆及序列分析

本研究以大青杨叶片总DNA为模板,用已登录的欧洲颤杨PtrCesA7保守序列设计引物,进行TD-PCR扩增并将得到的片段克隆到pMD18-T载体中。测序结果分析表明,片段长2341bp将该基因片段命名为PuCesA7。经BLASTX比对,此片段包含6个编码区,共1248bp,编码416个氨基酸。并与欧洲颤杨的PtrCesA7基因的同源性为98%,证明克隆的片段为纤维素合酶基因。将该片段与GenBank中登录的玉米(Zea mays)、拟南芥(Arabidopsis thaliana)、棉花(Gossypium hirsutum)、欧洲颤杨(Populus tremuloides)、新西兰辐射松(Pinups radiata D.Don)同源纤维素合酶基因序列比对并进行进化树分析。结果表明,大青杨纤维素合酶基因片段均与它们的亲缘关系较远。

Cloning and Sequence Analysis of Cellulose Synthase PuCesA 7 Fragment from Populus ussuriensis Kom.

In this study,we utilized total DNA of leaves from Populus ussuriensis Kom.to be as template for TD-PCR amplification and The obtained fragment were cloned into pMD 18-T vector by using specific primers,which designed based on conservative regions of PtrCesA 7 gene of Populus tremuloides.Sequence analysis showed that the cloned fragment was 2 341 bp,named as PuCesA 7.BLASTX results in the nucleotide sequences revealed that the gene fragment was 1 248 bp,containing six exons and encoding 416 amino acids.Comparison with the PircesA 7 gene from Populus tremuloides,the holomogy showed 98%.These results indicated that the fragment was cellulose synthase gene.Sequence alignment and phylogenetic analysis of PuCesA 7 gene among those of Zea mays,A rabidopsis thaliana,Gossypium hirsutum,Populus tremuloides and Pinups radiata D.Don from GenBank indicated that PuCesA 7 gene ofPopulus ussuriensis Kom.had a far relationship with the others.