The Nucleotide Sequence and Expression in E. coli of the Pieris rapae Granulosis Virus Granulin Gene

According to the Granulin gene’s 12 highly conservative nucleotides (TATAAGGAATTT) upstream from the protein intiation codon and the sequence of the Pieris brassiae granulovirus (PbGV) granulin gene, primers were designed to amplify the Pieris rapae granulosis virus (PrGV) granulin gene. The product of PCR amplification was about 850bp. The result of sequencing the segment showed the complete nucleotide sequence of the segment was 855bp. The segment contained one ORF encoding PrGV granulin gene which starts at position 38～40 and ends at position 779～781. The length of the ORF is 744bp and the length of predicted protein is 247 amino acid with MW of about 2.9178×104D. Comparison of the identities with other granulosis virus granulin gene showed the granulin gene is highly conserved in granulosis virus. The PrGV granulin gene was inserted into an exprssion vector, PET-28a, to yield the recombinant expression plasmid pet-28a-Gran. After induction by IPTG, the result of SDS-PAGE showed the expression of PrGV granulin in E. coli was obtained.