[Cloning of plasmid pBMB2062 in Bacillus thuringiensis strain YBT-1520 and construction of plasmid vector with genetic stability].
A small plasmid pBMB2062 in Bacillus thuringiensis strain YBT-1520 was cloned and sequenced. Its 2,062 bp sequence contains two potential open reading frames (orfs). The orf1 and orf2 encode a tentative replication initial protein consisting of 289 amino acid residues and a tentative replication protein consisting of 80 amino acid residues, respectively. Two homological plasmids were found by Blast searching. There are 23 nucleotides difference occurring among three of the plasmids. The difference occurred in the orf1 causes different encoding capability. Comparing with the orf1 in pBMB2062, the orf1 in the homological plasmids are truncated, one at the N-terminal and another at the C-terminal. cDNA synthesis and PCR detection showed that the mRNA corresponding to orf1 in pBMB2062 really occurs. Shuttle vectors were constructed based on pBMB2062 and showed the ability to express insecticidal crystal gene. Under nonselective condition, recombinant plasmids based on pBMB2062 were genetically stable.
Sun
M
M
College of Life Science and Technology, Huazhong Agricultural University, Key Laboratory of Agricultural Microbiology, Ministry of Agriculture, Wuhan, Hubei 430070, China.
Wei
F
F
Liu
Z D
ZD
Yu
Z N
ZN
D000595:Amino Acid Sequence; D001413:Bacillus thuringiensis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids