[日本血吸虫尾蚴cDNA文库的构建与鉴定]
[Construction and characterization of the cDNA library from Schistosoma japonicum cercariae].
作者信息
Chen Xiao-guang, Li Hua, Peng Hong-juan, Shen Shu-man, Fung Ming-chiu
机构信息
Department of Parasitology, Institute of Tropical Medicine, First Military Medical University, Guangzhou 510515.
出版信息
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2002;20(1):39-41.
OBJECTIVE
To construct a cDNA library from Schistosoma japonicum cercariae.
METHODS
Total RNA was extracted from 500,000 S. japonicum cercariae and used to synthesize the double-stranded cDNA and cloned into the TriplEx2 vector using "SMART cDNA Construction Kit".
RESULTS
The primary titer of the constructed cDNA library is 1.8 x 10(7) pfu/ml and the titer of amplified library is 2.5 x 10(10) pfu/ml. The average size of inserts is 1.075 kb. The recombinant efficiency is 94.4%. The full length cDNA of S. japonicum TPI and JF-2 genes was successfully amplified from the library.
CONCLUSION
A cDNA library of S. japonicum cercariae has been constructed successfully.
目的
构建日本血吸虫尾蚴的cDNA文库。
方法
从500,000条日本血吸虫尾蚴中提取总RNA，用于合成双链cDNA，并使用“SMART cDNA构建试剂盒”克隆到TriplEx2载体中。
结果
构建的cDNA文库的原始滴度为1.8×10⁷ pfu/ml，扩增文库的滴度为2.5×10¹⁰ pfu/ml。插入片段的平均大小为1.075 kb。重组效率为94.4%。从文库中成功扩增出日本血吸虫TPI和JF-2基因的全长cDNA。
结论
已成功构建日本血吸虫尾蚴的cDNA文库。
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