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THE CLONING, SEQUENCING AND EXPRESSION OF THE LdMNPV POLYHEDRIN GENE
Author(s) -
Tong LIN,
Chuanxi ZHANG,
Chengcai AN,
Zhiyini WANG,
Kuanyu LIU,
Huayi YUAN
Publication year - 2002
Publication title -
insect science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 45
eISSN - 1744-7917
pISSN - 1672-9609
DOI - 10.1111/j.1744-7917.2002.tb00171.x
Subject(s) - polyhedrin , open reading frame , gene , biology , cloning (programming) , genetics , expression vector , plasmid , nucleic acid sequence , molecular cloning , microbiology and biotechnology , peptide sequence , recombinant dna , spodoptera , computer science , programming language
LdMNPV ‐ NEFU isolate collected from the forestry farm of Northeast Forestry University was purified and the genomic DNA of MMNPV was extracted. The LdMNPV polyhedrin gene was cloned by PCR. The results showed that the sequence was an open reading frame (ORF) of 735bp capable of encoding 245 amino acids. The polyhedrin gene sequences of the MMNPV‐NEFU isolate and a Canada strain, MMNPV‐G differed in 5 bases. The polyhedrin gene of the LdMNPV‐NEFU isolate contained C, G, T, C and G at 54, 109,379, 508 and 701 sites from the start codon, but the LdMNPV‐G isolate contained G, C, C, T and T at the corresponding sites respectively. The same amino acids were encoded by the two ORF sequences, with the exception that Asp and His are encoded by GAC on the polyhedrin gene sequence of the LdMNPV‐NEFU isolate and by CAC in the MMNPV‐G isolate. The MMNPV polyhedrin gene was expressed in E. coli BL21 (DE3) by the pT7–7 plasmid vector.