Li Jie, Wang Zhi-gang, Hao Xi-yan, Chen Xian-wei, Wang Hong-xia, Li Shu-yu, Yang Jiao-fu, Yang Jun
College of Life Science, Inner Mongolia University, Key Laboratory of Mammal Reproductive Biology and Biotechnology, Ministry of Education, Hohhot 010021, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Apr;28(2):125-8.
To clone and express P-29 gene of Echinococcus granulosus, and analyze its immunoreactivity.
Total RNA was extracted from the hydatid cyst of E. granulosus and its P-29 gene was amplified by RT-PCR. The PCR product was cloned into pET44a(+) vector. The recombinant plasmid was transformed into E. coli BL21 (DE3) and followed by expression of the protein induced by IPTG. The protein was purified, and tested by SDS-PAGE. Its immunoreactivity was examined by Western blotting.
The recombinant expression plasmid was identified by PCR, double endonuclease digestion and sequencing. The recombinant Nus-P-29 was about Mr 93 000 with a concentration of 0.78 mg/ml. It was recognized by the sera of cystic echinococcosis patients.
The P-29 gene has been expressed with adequate immunoreactivity.
克隆并表达细粒棘球绦虫P-29基因，并分析其免疫反应性。
从细粒棘球绦虫的包虫囊肿中提取总RNA，通过RT-PCR扩增其P-29基因。将PCR产物克隆到pET44a(+)载体中。将重组质粒转化到大肠杆菌BL21(DE3)中，然后用IPTG诱导该蛋白表达。对该蛋白进行纯化，并通过SDS-PAGE进行检测。通过Western印迹法检测其免疫反应性。
通过PCR、双酶切和测序鉴定重组表达质粒。重组Nus-P-29的分子量约为93 000，浓度为0.78 mg/ml。它能被囊性棘球蚴病患者的血清识别。
P-29基因已成功表达，且具有足够的免疫反应性。
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