[间日疟原虫红细胞内期全长cDNA文库的构建]

[Construction of full-length cDNA library of erythrocytic stage Plasmodium vivax].

作者信息

Fang Qiang, Xia Hui, Yuan Yuan-Ying, Cao Jun, Wang Xue-Mei, Qi Wen-Juan, Gao Qi

机构信息

Department of Microbiology and Parasitology, Bengbu Medical College; Anhui Key Laboratory of Infection and Immunity, Bengbu 233030, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Jun 30;28(3):239-41.

PMID:20806516

Abstract

Blood samples were collected from vivax malaria patients without antimalarial drug therapy. After filtration through Plasmodipur filter to remove white blood cells, Plasmodium vivax-infected RBCs were enriched by Percoll. Total 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 x 10(6). The length of inserted fragment ranged from 900 to 2 500 bp, and the recombination efficiency accounted for 97.2%.

摘要

从未经抗疟药物治疗的间日疟患者采集血样。通过Plasmodipur过滤器过滤以去除白细胞后，用Percoll富集间日疟原虫感染的红细胞。分离红细胞中的总间日疟原虫。使用SMART cDNA文库构建试剂盒构建间日疟原虫红细胞期的全长cDNA文库。评估文库的体积和重组率。通过PCR扩增鉴定插入片段。cDNA文库的滴度为1.14×10(6)。插入片段长度为900至2500 bp，重组效率为97.2%。