摘要
TTW4是本实验室自内蒙古盐湖中分离得到的一株中度嗜盐菌,经形态学、生理生化和16srDNA序列分析确定其属于Halomonas属。通过紫外诱变获得该菌株具有利福平抗性的突变株UV-1。利用三亲本杂交得到UV-1的8株盐敏感突变株。碘结晶法测定UV-1和突变株胞内甜菜碱含量,UV-1胞内甜菜碱最多可达190μg/mg,盐敏感突变株胞内甜菜碱含量有明显降低,一般在150μg/mg以下,最低的只有110μg/mg。以其中一株盐敏感突变株S3为实验材料,反向PCR扩增转座子Tn5插入位点侧翼序列,BLAST分析表明插入位点为色氨酸合酶α亚基基因,推测该基因与耐盐功能相关。
TTW4 is a moderate halophilic bacteria strain isolated from the salt lake in Inner Mongolia China.TTW4 is identified as Halomonas.We got the rifampicin resistant mutant strain UV-1 through ultraviolet.We got 8 strains of salt sensitive mutant strains from UV-1 by triparental mating.Iodine crystallizing method determination of betaine content of the bacterial cell.UV-1 can accumulate the betaine reach a maximum of 190μg/mg.Salt sensitive mutant strains of intracellular accumulation of betaine content have significantly lower,usually under 150μg/mg,the lowest is only 110μg/mg.Let S3 as a salttolerant genes cloning experiment material which salt tolerance ability significantly lower.Inverse PCR amplification transposon Tn5 insertional locus flanking sequence.BLAST analysis indicates that the insertion site is tryptophan synthase alpha subunit gene,presumably the gene related to salt tolerance function.
出处
《青岛农业大学学报（自然科学版）》
2013年第3期204-210,共7页
Journal of Qingdao Agricultural University(Natural Science)
基金
国家自然基金(63070047)资助
关键词
中度嗜盐菌
盐敏感突变株
甜菜碱
耐盐基因
Moderately halophilic bacterium
Salt sensitive mutant
Betaine
Salt tolerant gene
