Cloning Expression and Characterization of Two Cysteine Proteinases of Clonorchis sinensis
- VernacularTitle:两个华支睾吸虫半胱氨酸蛋白酶的基因克隆、表达与定性
- Author:
Fuquan PEI
;
Nagano ISAO
;
Jun WU
;
Zhiliang WU
;
Huier CUI
;
Takahashi YUZO
;
Bo PAN
;
Yueyi FANG
- Publication Type:Journal Article
- Keywords:
Clonorchis sinensis;
recombinant cysteine proteinases;
expression;
characterization
- From:
Journal of Tropical Medicine
2004;4(1):10-14,55
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express the valuable Clonorchis sinensis antigen molecules which can be applied to the diagnosis of clonorchiasis. Methods Based on the sequences (Genbank) No. AF271091 (CysA) and No.AF093242 (CysB), primers were designed to amplify the two C. sinensis cysteine proteinase genes and expressed in E.cloi. The expressed proteins were purified by affinity chromatography and then tested for their immunological characters.Results The two genes were successfully cloned and expressed. Western blot showed that CysB had strong reaction with clonorchiasis sera and very weak reaction with schistosomiasis sera, while CysA showed no reactivity with the probed sera. Immunohistochemistry showed that both proteins were mainly located in adult worm intestines and the intrauterine eggs.Conclusions The results suggested that, of the two expressed C. sinensis proteins, CysB had good antigenic reactivity against sera from patients. It is a potential candidate of diagnostic antigens for clonorchiasis.
