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Full-Length cDNA Cloning and Bioinformatic Analysis of Schistosoma Japonicum Infection Resistance Associated Gene E77.43 in Microtus Fortis

Author: XiongDeHui
Tutor: HuWeiXin
School: Central South University
Course: Biochemistry and Molecular Biology
Keywords: Microtus fortis Schistosoma japonicum resistance associated gene RACE
CLC: Q78
Type: Master's thesis
Year: 2009
Downloads: 48
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Abstract


Schistosomiasis is a zoonoses.Epidemiological survey and experimental research of Microtus fortis infected with schistosoma japonicum cercariae suggested that Microtus fortis exhibits innate resistance to schistosoma japonicum infection,furthermore,the innate resistance to schistosoma japonicum in Microtus fortis inherit stably.In previous study,we constructed bone marrow cDNA library of Microtus Fortis’s,isolated and characterized E77.43 clone from the library by expression cloning techniques.The transient expression products of E77.43 can inhibit schistosomula japonicum activity obviously in vitro.To obtain the full-length cDNA of E77.43,We harvested the bone marrow cells of Microtus Fortis,extracted the total RNA,constructed a cDNA library.3’and 5’end of E77.43 were amplified by RACE and homologous genes amplification techniques separately, and then sequenced.After obtained the 3’and 5’ends of E77.43,we aligned the E77.43 sequences,the full-length cDNA of E77.43 is 3274 bp. Bioinformatics analysis show the opening reading frame of E77.43 encodes 392 amino acid,the molecular weight is 42.8 kDa,isoelectric point is 6.07.E77.43 coding protein was supposed to be an acidic protein in nuclear.We found the coding product of E77.43 could be integrin-linked kinase-associated serine/threonine phosphatase,which play a key role in regulation of cell survival and apoptosis,the interactive protein with it is integrin-linked protein kinase.The obtaining of the full-length cDNA of E77.43 laid a strong basis for the further research.

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