摘要
根据GenBank报道的谷氨酸棒杆菌ATCC13032序列,设计并合成一对引物,获得了全长argB基因片段.将其克隆到pGEM T Easy载体中,经测序鉴定后,以其为DIG标记探针,通过SouthernBlot分析钝齿棒杆菌A.S1.542与谷氨酸棒杆菌ATCC13032,argB基因核苷酸同源性高达99%,氨基酸序列95%相同.
The argB sequence was obtained by the designing and synthesizing a pair of primer,based on the report of GenBank about the sequence of Corynebacterium glutamicum ATCC 13032. The argB gene was cloned into plasmid pGEM-T-Easy. The recombinant strain harboring the pGEM-argB plasmid was verified by PCR reaction, plasmid extraction with alkaline method, digestion with restriction endonuclease BamHI and EcoRI, and DNA sequencing. The result of Southern blot indicated that the homology of argB between C.crenatum A.S1.542 and C.glutamcium ATCC13032 was more than 90% by the DIG-argB probe to screen the genomic libraries. The nucleotide sequence and the predicted amino acid sequence of argB from C.crenatum were 99% and 100% identical with those of the argB from C.glutamicum.
基金
江苏省"十五"攻关项目(编号:BE20011041)资助课题.
