[日本血吸虫膜蛋白单链抗体（scFv）的基因克隆、构建及表达]
[Gene cloning, construction and expression of single-chain Fv (scFv) against the membrane protein of Schistosoma japonicum].
作者信息
Yu X C, Jiang X, Huang H M, Zhang Z, Lin Q, Guan X H, Huang H L
机构信息
Institute of Genetics, Chinese Academy of Sciences, Beijing 100101.
出版信息
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2001;19(3):135-40.
OBJECTIVE
To construct single chain antibody specific to membrane protein of Schistosoma japonicum by genetic engineering technique.
METHODS
The VH (heavy-chain variable region) and VL(light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using VH and VL genes, then expressed by IPTG.
RESULTS
The VH and VL genes were obtained through PCR. The DNA sequences showed that VH and VL were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36.2 kDa.
CONCLUSION
A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.
目的
通过基因工程技术构建针对日本血吸虫膜蛋白的单链抗体。
方法
从NP11 - 4细胞系基因组DNA中通过PCR扩增重链可变区（VH）和轻链可变区（VL）基因，并用桑格法进行测序。利用VH和VL基因在pTHA90载体中构建单链抗体片段（ScFv），然后用异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达。
结果
通过PCR获得了VH和VL基因。DNA序列显示VH和VL是新的抗体可变区基因，并已在GenBank注册。在pTHA90载体中成功构建了带有（Gly4Ser）3间隔肽的ScFv基因。ScFv表达为约36.2 kDa的硫氧还蛋白融合蛋白。
结论
构建并表达了针对日本血吸虫膜蛋白的特异性ScFv。
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