摘要
传统筛种方法具有随机性、盲目性,而PCR筛菌,增加了目的性和效率。在基因序列数据库中查找益生双歧杆菌的16S rDNA共有序列,找出它们共有的特异性序列,设计了PCR引物。平板培养分离,初步筛选双歧杆菌菌落。根据细菌16S rDNA和23S rDNA两侧高度保守区域设计通用引物,提取菌落基因组DNA,扩增16S-23S rDNA间区序列,并送测序。通过序列同源性比对分析,鉴定、筛选到人体长双歧杆菌。
It increased the purpose and efficiency using PCR to screening bacteria,while traditional methods have the characters of randomness and blindness. Primers for PCR were designed based on nucleotide sequences of the common and specific region of 16S rDNA for the probi- otic Bifidobacterium strains .After culturing on the agar plate,the colonies were primarily screened with PCR. A pair of universal primers were designed according to the highly conserved regions of 16S rDNA and 23S rDNA.After extraction of genomic DNA from the screened colonies,the PCR products of 16S-23S rDNA intergenic spacer were sequenced. The isolated colonies were identified according to sequence analysis using Blast in the Genbank.At last, the strains of Bifidobacterium longum have been successfully screened out.
出处
《中国乳品工业》
CAS
北大核心
2012年第12期26-27,47,共3页
China Dairy Industry
基金
乳业生物技术国家重点实验室筹建项目(10dz2221100)
上海乳业生物工程技术研究中心项目(09DZ2251400)
关键词
长双歧杆菌
PCR
定向筛选
菌种鉴定
Bifidobacterium longum
PCR
directional screening
strain identification
