一株高产脂肪酶产生菌16S rDNA的序列分析

目的]对分离获得的高产脂肪酶菌株进行鉴定,为其改造和更好利用奠定基础.方法]对从食堂下水道中分离获得的一株高效产脂肪酶细菌(JLΠ-4)进行培养,提取其基因组DNA.设计16S rDNA通用引物,扩增16S rDNA基因片段,并连接到pUC19-T载体上,转化大肠杆菌DH5X,经PCR鉴定的阳性克隆摇菌培养后测序.结果]提取获得较高质量的基因组DNA,扩增获得新分离菌株16S rDNA基因片段,长度为1528 bp,BLAST相似性比对分析结果表明,其与伯克霍尔德氏菌16S rDNA序列相似性达97%,是一株与伯克霍尔德氏菌最近的革兰氏阴性菌.结论]初步将高产脂肪酶细菌JTΠ-4鉴定为唐菖蒲伯克霍尔德菌.

Sequence analysis of 16S rDNA derived from a high yield lipase strain

Objective]The present study was conducted to identify the separated high yield lipase strain to provide theoretical research references and to enhance the high yield lipase strain’s transformation and application.Method]A strain of bacterium with high yield lipase JLП-4 was isolated from the sewage of a canteen,then its genomic DNA was extracted.The gene fragments of 16S rDNA were amplified using 16S rDNA universal primers and connected to pUC19-T vector;the fragments are then transformed into E.coli DH5X.The positive clones identified by the PCR method were cultured and sequenced.Result]Quality genome DNA was successfully extracted.The 16S rDNA gene fragments of newly isolated strain were amplified with the length of 1528 bp.According to comparison analysis of BLAST,16S rDNA sequence similarity between the strains and Burkholderia(DQ355168) were 97%,so the lipase producing strains were identified as gram-negative bacteria that were most similar to the structures of Burkholderia.Conclusion]The high-yield lipase JLП-4 was primarily identified as Burkholderia gladioli.