Identification, expression and in situ hybridization of an eggshell protein gene from Fasciola hepatica.
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.
Zurita
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M
Department of Pharmacology, Stanford University School of Medicine, CA 94305.
Bieber
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D
Mansour
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D000818:Animals; D003001:Cloning, Molecular; D004247:DNA; D004527:Egg Proteins / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005210:Fasciola hepatica / Q000235:genetics* / Q000648:ultrastructure; D015870:Gene Expression; D008970:Molecular Weight; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000032:analysis; D001616:beta-Galactosidase
D004527:Egg Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA; D001616:beta-Galactosidase