短小芽孢杆菌肉碱转运系统opuC基因的克隆与序列分析

摘要

以前期获得的ω_(-1)-羟基脂肪酸高产突变菌株短小芽孢杆菌(Bacillus pumilus)M-F641的总DNA为模板,利用Primer Premier 5.0软件设计4对引物,对决定长链脂肪酸无效降解途径中肉碱转运的OpuC转运系统的基因进行克隆,成功获得了opuCA、opuCB、opuCC和opuCD的基因序列,并利用MEGA 3.1、DNAStar等软件进行序列分析.研究内容将为进一步利用短小芽孢杆菌长链脂肪酸高效转化生产ω_(-1)-羟基脂肪酸菌株奠定基础.%Chromosomal DNA of ω_(-1)-hydroxy fatty acids(HFA)high-yielding mutant Bacillus pumilus strains M-F641 as a template, the gene of OpuC transport system deciding carnitine transport in the course of long-chain fatty acid degradation was amplified by PCR,by which the sequences of opuC gene and the amino acid sequences of OpuC transport system were obtained. Results showed that the length of the opuCA, opuCB, opuCC and opuCD sequences are 1 162 bp,654 bp,951 bp and 690 bp, respectively. By using the MEGA 3. 1 DNAStar software for sequence analysis as well as phylogenetic analysis of OPUC,the results of which showed that the sequence of the fragment from B. pumilus M-F641 had a high similarity with the OPUC transport system of B. pumilus SAFR-032 reported by the reference. This research could provide the base for bioconversion of long-chain fatty acids for ω_(-1)-HFA production with high efficiency by Bacillus pumilus.