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肝片吸虫卵壳蛋白基因的鉴定、表达及原位杂交

Identification, expression and in situ hybridization of an eggshell protein gene from Fasciola hepatica.

作者信息

Zurita M, Bieber D, Mansour T E

机构信息

Department of Pharmacology, Stanford University School of Medicine, CA 94305.

出版信息

Mol Biochem Parasitol. 1989 Nov;37(1):11-7. doi: 10.1016/0166-6851(89)90097-2.

Abstract

The molecular basis of egg formation in the parasitic liver fluke, Fasciola hepatica, was investigated by isolating and characterizing an abundant cDNA from a female genital complex cDNA library. It was expressed in Escherichia coli as a beta-galactosidase fusion protein, which was purified and used to produce polyclonal antibodies. Using immunoblots, the antiserum recognized two soluble constituents of isolated egg shells, both significantly larger than predicted from cDNA sequencing. Using in situ hybridization, the message was detected in cells in the adult vitelline follicles. Eggshell protein mRNA expressed in E. coli will provide a source of precursor protein for further studies of parasite eggshell formation.

摘要

通过从雌性生殖复合体cDNA文库中分离并鉴定一种丰富的cDNA,对寄生性肝吸虫(肝片吸虫)卵子形成的分子基础进行了研究。它在大肠杆菌中作为β-半乳糖苷酶融合蛋白表达,该融合蛋白经纯化后用于制备多克隆抗体。利用免疫印迹法,抗血清识别出分离出的卵壳的两种可溶性成分,二者均明显大于cDNA测序预测的大小。利用原位杂交技术,在成年卵黄滤泡的细胞中检测到了该信息。在大肠杆菌中表达的卵壳蛋白mRNA将为进一步研究寄生虫卵壳形成提供前体蛋白来源。

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