Cloning, expression and sequence analysis of autophagy-related protein 3(APG3-like)gene from Taenia asginata asiatica

Objective To search and identify novel gene by screening the cDNA library of adult Taeniasa ginata a siatica and to insert the screened gene into autophagy-related prote in 3(APG 3),then to analyze and predict the structure and characteristics of Ta.APG3 gene by bio-informatics.Methods By the bioinformatics analyzing tools and combining other complicated bio informatics software package,the characte ristics of the prote in was analyzed.The gene was inserted into the prokaryo tic expression vectors pET-28a(+),then was amplified with PCR and endonuc lease digestion and then expressed with IPTG induction.The recombinant prote in was detected by SDS-PAG E.Results The novel cDN A sequence encoding Ta.APG 3 has a molecular mass of 37498.5,and anisoe lectric point of 4171.T he gene is 1331bp coding 332 amino acids.PCR,double enzyme digestion and DNA sequencing all indicated that pET-28a(+)and APG 3 recom binant plasmid was successfully constructed.SDS-PAG E results showed that the gene expression took place in Escherichia coli BL 21/D E3,and highly pureprote in was achieved after urea inclusion body deposits.Conclusion Anovel gene of Taenia saginata a siatica was constructed in prokary otic expression vector and its fusion prote in was expre ssed successfully.