CAJ | 학술논문

目的克隆间日疟原虫河南分离株与湖北分离株红内期18S rDNA,并进行同源性分析.方法采用PCR方法从间日疟患者血样DNA中扩增间日疟原虫18S rDNA,纯化后与pGEM-Teasy质粒连接,转化大肠埃希氏菌JM109;阳性克隆质粒经双酶切鉴定后,进行序列测定,采用BLAST和MEGA4生物软件分析同源性. 结果间日疟原虫18S rDNA扩增片段大小为998 bp;阳性克隆重组质粒经双酶切鉴定,与预期结果相符;序列测定结果显示,河南、湖北2分离株间日疟原虫18S rDNA序列完全相同,与GenBank中报道的12株间日疟原虫相同序列进行比对,其同源性均大于99％;用邻位连接法(neigh-bor-joining,NJ)和非加权组平均法(UPGMA)2种方法构建系统发生树发现,河南分离株、湖北分离株与间日疟原虫X13926.1株遗传距离小,同属一个分支.结论克隆了间日疟原虫河南与湖北分离株红内期18S rDNA,该基因序列在不同地理株间遗传稳定.
목적 극륭간일학원충하남분리주여호북분리주홍내기18S rDNA,병진행동원성분석.방법 채용PCR방법종간일학환자혈양DNA중확증간일학원충18S rDNA,순화후여pGEM-Teasy질립련접,전화대장애희씨균JM109;양성극륭질립경쌍매절감정후,진행서렬측정,채용BLAST화MEGA4생물연건분석동원성. 결과 간일학원충18S rDNA확증편단대소위998 bp;양성극륭중조질립경쌍매절감정,여예기결과상부;서렬측정결과현시,하남、호북2분리주간일학원충18S rDNA서렬완전상동,여GenBank중보도적12주간일학원충상동서렬진행비대,기동원성균대우99％;용린위련접법(neigh-bor-joining,NJ)화비가권조평균법(UPGMA)2충방법구건계통발생수발현,하남분리주、호북분리주여간일학원충X13926.1주유전거리소,동속일개분지.결론 극륭료간일학원충하남여호북분리주홍내기18S rDNA,해기인서렬재불동지리주간유전은정.
Objective To clone and homology analyze the sequences of blood stage 18S rRNA-encoding gene fragment of two P.vivax isolates from Henan and Hubei provinces in China.Methods The 18S rDNA fragments were amplified by PCR from the DNA extracted from two P.vivax infection blood samples.After purification,the gene fragments were ligated with plasmid pGEM-Teasy to construct recombinant plasmids,and transformed into E.coli JM109.Positive clones were identified by double enzymes digestion methods.The sequences of inserted 18S rDNA fragments were finally determined and analyzed with BLAST and MEGA4 biological software.Results The amplified 18S rDNA fragments of two isolates were about 998 bp in length,and the 18S rDNA sequence of Henan isolate was same as that of Hubei isolate.As aligned with the corresponding sequences of twelve P.vivax strains deposited in the GenBank database,the indentity of nucleotides was more than 99％ respectively.Based on the 18S rDNA sequence,phylogenetic analysis with neighbor-joining and UPGMA methods indicated that two isolates have close association with P.vivax X13926.1 strain.Conclusion The blood stage 18S rDNA fragments of two P.vivax isolates were cloned,which were relatively conserved among different P.vivax isolates.