构建间日疟原虫红内期全长cDNA文库  

Construction of Full-length cDNA Library of Erythrocytic Stage Plasmodium vivax

作　　者：方强[1] 夏惠[1] 袁远英[1] 曹俊[2] 王雪梅[1] 齐文娟[1] 高琪[2] 

机构地区：[1]蚌埠医学院病原生物学教研室,安徽省感染与免疫重点实验室,蚌埠233030 [2]江苏省寄生虫病防治研究所,卫生部寄生虫病预防与控制技术重点实验室,无锡214064

出　　处：《中国寄生虫学与寄生虫病杂志》2010年第3期239-240,F0003,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家传染病科技重大专项(No.2008ZX10004-011);卫生部寄生虫病预防与控制技术重点实验室开放课题(No.WK008-002)~~

摘　　要：采集未经治疗的间日疟患者血样,滤去白细胞后浓集含虫红细胞,提取总RNA,用SMARTcDNA文库构建试剂盒构建间日疟原虫红内期全长cDNA文库。测定cDNA文库库容,蓝白斑筛选,计算重组率。随机挑选48个菌落,用M13+/-引物进行PCR鉴定插入片段大小。构建的间日疟原虫红内期全长cDNA文库原始库容为1.14×106,重组率为97.2%,重组子插入cDNA片段大小为900～2500bp。Blood samples were collected from vivax malaria patients without antimalarial drug therapy. After filtra-tion through Plasmodipur filter to remove white blood cells, Plasmodium vivax-infected RBCs were enriched by Percoll. Total RNA of P. vivax in red blood cells was isolated. A full-length cDNA library of erythrocytic stage P. vivax was constructed by the SMART cDNA library construction kit. The volume and recombinant rate of the library were evaluated. The inserted fragments were identified by PCR amplification. The titer of cDNA library was 1.14×106. The length of inserted fragment ranged from 900 to 2 500 bp, and the recombination efficiency accounted for 97.2%.

关 键 词：间日疟原虫 红内期 CDNA文库 

分 类 号：R91[医药卫生—药学]