摘要
分离并鉴定了长链烷烃降解菌Pseudomonasaeruginosa1785和P.marginata766烃羟化酶基因alkB片段.根据烃羟化酶的保守氨基酸序列,设计兼并引物,扩增P.aeruginosa1785和P.marginata766的alkB片段,获得了目标产物.经DNA测序和氨基酸序列分析,证实目标片段编码的肽段含有烃羟化酶的特征基序.由此确认采用该方法分离到了长链烷烃降解基因的alkB同源体片段.DNA序列比对结果表明,P.aeruginosa1785和P.marginata766的alkB片段与P.aeruginosaPAO1的alkB1和alkB2的相似性分别达到95.7%和94.8%.这些alkB片段可用于分析烃降解微生物群落结构.
alkB fragments of two long-chain alkane degrading bacterium strains, Pseudomonas aeruginosa 1785 and P. marginata 766 were isolated and identified. Highly degenerate primers were designed for PCR amplification of alkB homology in P. aeruginosa 1785 and P. marginata 766 based on a number of highly conserved sequence motifs. From DNA sequencing and analysis of peptide sequence, signature motifs of alkane hydroxylase were found in target product of PCR. Sequence alignment showed that the alkB fragment from P. aeruginosa 1785 and P. marginata 766 had 95.7% and 94.8% sequence identity to correspond fragment of alkB1 and alkB2 in P. aeruginosa PAO1, respectively. These alkB fragments would be very. useful as molecular probes for detecting function groups in analyses of microbial community structure relating to alkane degrading bacteria.
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2006年第4期562-565,共4页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金(No.50374038)
高等学校博士点基金(No.20030251002)
教育部重点项目(No.03071)
上海市科委资助项目(No.045407017)~~
关键词
长链烷烃降解菌
alkB片段
PCR扩增
long-chain alkane degrading bacterium
alkB fragment
PCR amplification