食品科学 ›› 2017, Vol. 38 ›› Issue (24): 8-14.doi: 10.7506/spkx1002-6630-201724002

• 生物工程 • 上一篇    下一篇

婴儿粪便长双歧杆菌的分离与多样性分析

张秋雪,刘晓婵,朱宗涛,万峰,孙思睿,贾芳芳,孟祥晨   

  1. (东北农业大学 乳品科学教育部重点实验室,黑龙江?哈尔滨 150030)
  • 出版日期:2017-12-25 发布日期:2017-12-07
  • 基金资助:
    国家自然科学基金面上项目(31671917)

Isolation and Diversity Analysis of Bifidobacterium longum from Infant Faeces

ZHANG Qiuxue, LIU Xiaochan, ZHU Zongtao, WAN Feng, SUN Sirui, JIA Fangfang, MENG Xiangchen   

  1. (Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China)
  • Online:2017-12-25 Published:2017-12-07

摘要: 本研究旨在分析婴儿粪便长双歧杆菌的多样性,采用改良MRS培养基从哈尔滨地区健康婴儿粪便中分离双歧杆菌,采用生理生化实验结合16S rDNA和热应激蛋白60基因(heat-shock protein 60,hsp60)同源性分析鉴定分离株,利用同源性分析及随机扩增多态性DNA(random amplified polymorphism DNA,RAPD)、多位点序列分型(multilocus sequence typing,MLST)技术进一步分析长双歧杆菌多态性。实验从11 个婴儿体内分离得到18 株厌氧的细菌菌株,经形态学分析、生理生化实验、16S rDNA测序及hsp60测序实验发现,其中7 株为长双歧杆菌长亚种,3?株为长双歧杆菌婴儿亚种。RAPD和MLST结果表明:7?株长双歧杆菌长亚种为6?个基因型,3?株长双歧杆菌婴儿亚种为2?个基因型,上述结果说明不同人源婴儿粪便长双歧杆菌基因型差异较大。

关键词: 长双歧杆菌, 热应激蛋白60基因(hsp60), 随机扩增多态性DNA(RAPD), 多位点序列分型(MLST)

Abstract: The aim of this experiment was to study the diversity of Bifidobacterium longum isolated from infant feces. Using modified MRS medium, bifidobacterial strains were isolated from the feces of healthy infants in Harbin. To identify and analyze the diversity of B. longum strains, we used physiological and biochemical experiments, 16S rDNA homology analysis, heat-shock protein 60 gene (hsp60) homology analysis, random amplified polymorphism DNA (RAPD) and multilocus sequence typing (MLST). Eighteen anaerobic strains were isolated, among which, 7 were identified as B. longum subsp. longum, and 3 as B. longum subsp. infantis according to morphology, physical and biochemical properties, 16S rDNA homology analysis and hsp60 homology analysis. The results of RAPD and MLST showed that the 7 strains of B. longum subsp. longum belonged to 6 genotypes while the 3 strains of B. longum subsp. infantis belonged to 2 genotypes. Overall, these results showed that there were differences in the diversity of B. longum isolated from infant feces.

Key words: Bifidobacterium longum, heat-shock protein 60 gene (hsp60), random amplified polymorphism DNA (RAPD), multilocus sequence typing (MLST)

中图分类号: