畜牧与饲料科学 ›› 2013, Vol. 34 ›› Issue (5): 1-1.doi: 10.12160/j.issn.1672-5190.2013.05.002

• 基础科学 • 上一篇    下一篇

瘤胃表皮葡萄球菌的分离鉴定及其β-葡聚糖酶基因的克隆和在乳酸球菌中表达的研究

呼和 苏少锋 刘永志 刘红葵 王蕴华   

  1. 内蒙古农牧业科学院,内蒙古呼和浩特010031
  • 出版日期:2013-05-20 发布日期:2013-05-20
  • 通讯作者: 呼和
  • 作者简介:呼和(1962-),男,研究员,博士,主要研究方向为饲料生物技术。

Identification of a Staphylococcus epidermidis Isolated from Rumen and the Cloning of β-glucanase Gene and Its Expression in Lactococcus lactis

HU He, SU Shao-feng, LIU Yong-zhi, LIU Hong-kui, WANG Yun-hua (Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot 010031, China)   

  • Online:2013-05-20 Published:2013-05-20

摘要: 为将瘤胃微生物纤维降解酶基因转化到乳酸菌中,从绵羊瘤胃内容物中分离到1株能够降解羧甲基纤维素钠的细菌,经过形态特征及细菌16SrDNA序列鉴定为表皮葡萄球菌。根据表皮葡萄球菌在GenBank中公示的β-葡聚糖酶基因序列设计PCR扩增引物,克隆到2个β-葡聚糖酶基因序列。将2个β-葡聚糖酶基因序列与载体pMG36e连接,构建β-葡聚糖酶基因表达载体,转化到大肠杆菌,并进一步转化到乳酸球菌MG1363,均能表达β-葡聚糖酶,表达酶活达1.47U/mL和1.54U/mL,比原表皮葡萄球菌酶活0.99U/mL高出1.49倍和1.56倍。

Abstract: In order to clone and express rumen microbial fibrolytic enzyme genes in a strain of Lactococcus lactis, a cellulose degrade bacterium from the rumen of the sheep was isolated by using improved CMC medium based on widely screen. The bacterium was identified as Staphylococcus epidermidis by using analysis of morphology and the conserved sequence 16S rDNA. Two β-glucanase genes were cloned from the bacterium chromosome DNA by original strain of Staphylococcus epidermidis. The two β-glucanase genes were connected with vector pMG36e to construct β-glucanase gene expression vector, and then transformed into E. coli and Lactococcus lactis MG1363. Both them could express β-glucanase, and their enzyme activities reached 1.47 U/mL and 1.54 U/mL respectively, which were higher 1.49 times and 1.56 times than that of original strain of Staphylococcus epidermidis (0.99 U/mL).

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