摘要
为了对食源性单核细胞增生型李斯特菌LM5567的arcD基因进行克隆和序列分析,实验利用PCR方法对arcD基因进行扩增,连接至pMD19-T载体,将连接产物转化至DH5接感受态细胞,筛选阳性菌株进行测序比对分析。结果显示:扩增获得的arcD基因序列长为915bp,克隆得到arcD基因的阳性转化子;生物信息学分析表明:arcD蛋白属于跨膜蛋白,无信号肽序列;LM5567的arcD基因核苷酸序列与02-6680株(4b,奶酪,加拿大)相似性99.9%,与10-0811株(1/2b,螃蟹,加拿大)相似性99.7%,与10-0809株(4b,粪便,加拿大)、81-0558株(4b,脑脊髓液,加拿大)、02-1103株、02-1792(4b,奶酪,加拿大)和81-0861株(4b,卷心菜,加拿大)相似性均为100%。LM5567arcD基因编码的氨基酸序列与上述菌株相似性为89.4%~89.9%。本实验成功克隆了arcD基因,为进一步探究arcD基因的功能奠定了基础。
To clone and analyse the arcD gene of Listeria monocytogenes LM5567. The arcD gene was amplified by PCR and ligated with pMD19-T vector.The ligation product was transformed into DH5 competent cells, and the positive strains were screened for sequencing analysis. The results showed that the length of the obtained arcD gene was 915 bp, and the positive transformants of the arcD gene were cloned. The positive strains were screened for sequencing analysis. The bioinformatics analysis showed that the arcD protein belongs to the transmembrane protein and has no signal peptide sequence. The nucleotide sequence of the arcD gene of LM5567 and 02-6680 strain(4 b, cheese, Canada) similarity 99.9%, similarity with 10-0811 strain(1/2 b, crab, Canada) 99.7%, with 10-0809 strain(4 b, feces, Canada), 81-The similarities of 0558 strains(4 b, cerebrospinal fluid, Canada), 02-1103 strains, 02-1792(4 b, cheese, Canada) and 81-0861 strains(4 b, cabbage, Canada) were 100%. The amino acid sequence encoded by the LM5567 arcD gene has a similarity to the above strain of 89.4% to 89.9%. This experiment successfully cloned the arcD gene, which laid the foundation for further exploration of the function of the arcD gene.
作者
刘扬扬
马勋
康立超
李红欢
钱凌霄
江婉琳
阮婷玉
LIU Yang-yang(College of Animal Science and Technology of Shihezi University,Shihezi 832003)
出处
《青海畜牧兽医杂志》
2019年第6期21-26,共6页
Chinese Qinghai Journal of Animal and Veterinary Sciences
基金
国家自然科学基金项目(31860712)
