摘要
目的查找9种细菌和1种真菌的水通道蛋白基因,为进一步探讨细菌水通道蛋白的生理功能奠定基础。方法根据细菌水通道蛋白的保守氨基酸序列,设计合成简并核苷酸引物。提取目标菌DNA,简并PCR扩增细菌水通道蛋白基因。扩增产物回收纯化,DNA测序,推导其氨基酸序列并BLAST分析。结果10种目标菌中,大肠埃希菌和痢疾志贺菌扩增出所需的DNA片段。电泳分析其分子质量约为400 bp和370 bp,其序列与细菌水通道蛋白-甘油易化蛋白基因高度同源,相似性分别为97%和94%;推导氨基酸序列与细菌水通道蛋白氨基酸高度同源,相似性为83%和87%。结论利用简并PCR在大肠埃希菌中扩增出甘油易化蛋白基因,并发现痢疾志贺菌中存在甘油易化蛋白基因。
Objective The genes of 9 bacteria and 1 fungus were examined to provide the basic foundations to determine the physiological functions of aquaporins.Methods In accordance with the conserved amino acid sequences of aquaporins in bacteria,degenerate nucleotide primers were designed and synthesized.DNA of objective bacteria was extracted and the genes of bacterial aquaporin were amplified by degenerate PCR.The amplified products were analyzed by gel electrophoresis.The amplified DNA was recovered and extracted, and then the amino acid sequences of the DNA were deduced and analyzed with BLAST. Results The DNA fragments needed were amplified from E. coli and S. dysenteriae among the 10 target bacteria. Eleetrophoretie analysis indicated that they were about 400 bp and 370 bp DNA, respectively. Their sequences were extremely homologous with Glp-F of bacterial aquaporin and their similarity was 97 % and 94 %, respectively. The amino acid sequences were extremely homologous with Glp-F and their similarity was 83% and 87%. Conclusion GIp-F genes were amplified from E. coli and S. dysenteriae by degenerate PCR
出处
《中国病原生物学杂志》
CSCD
2009年第11期818-822,共5页
Journal of Pathogen Biology
