斯达氏油脂酵母肌动蛋白基因克隆及序列分析  

Cloning and Sequence Analysis of Actin Gene of Oleaginous Yeast Lipomyces starkeyi

作　　者：唐伟[1,2] 谭海东[1] 张素芳[1] 赵宗保[1] 

机构地区：[1]中国科学院大连化学物理研究所,大连116023 [2]中国科学院研究生院,北京100049

出　　处：《生物技术通报》2009年第S1期280-283,299,共5页Biotechnology Bulletin

基　　金：国家863计划(2007AA05Z403);中国科学院知识创新工程重要方向项目(KGCX2-YW-336)

摘　　要：以产油真菌斯达氏油脂酵母(Lipomyces starkeyi AS 2.1560)为材料,提取总RNA,反转录成cDNA。根据真菌肌动蛋白(actin)保守核酸序列设计引物,PCR扩增后获得部分序列,再采用cDNA末端快速扩增技术获得了开放阅读框全长为1128 bp的cDNA序列,该序列编码蛋白质含375个氨基酸残基,等电点为5.49。同源性比较发现该基因(Accession No. EU258762)与其它已知真菌的actin基因相似性在75%以上,氨基酸序列相似性在90%以上。通过不同物种肌动蛋白进化树分析发现L.starkeyi与亚罗解脂酵母(Yarrowia lipolytica)有较近的素缘关系。该研究对探索L.starkeyi产油机制有一定的参考价值。Total RNA was extracted from the oleaginous yeast Lipomyces starkeyi and reversed transcripted to the first-strand cDNA. Primers were designed according to conserved sequence of actin gene from other microorganisms and the actin gene of L.starkeyi AS 2.1560(Lsactin) was isolated using methods of RT-PCR and rapid amplification of cDNA ends.The open reading frame of Lsatcin contains 1128 bp and encodes 375 amino acid residues with an isoelectric point of 5.49.When analyzed using the program BLAST,the DNA sequence of Lsactin(Accession No.EU258762) presents more than 75%identity scores with those of other fungi,while the protein sequence has more than 90%identity.Phylogenetic analyses of the evolutionary relationship revealed that Lsactin has a closer relationship with that from Y.lipolytica.This work provided useful information for further study on the molecular mechanism of lipid accumulation by L.starkeyi.

关 键 词：斯达氏油脂酵母 肌动蛋白 RNA提取 RACE 系统进化树 

分 类 号：Q78[生物学—分子生物学]