Genomic segments cloning and analysis of Cotesia plutellae polydnavirus using plasmid capture system.
Cotesia plutellae polydnaviruses (CpBV) has a segmented genome consisting of multiple circular double stranded DNAs. Recently, we have developed an easy, simple, and convenient system based on Tn7 transposition in order to clone genomic segments of CpBV in Escherichia coli cell and designated plasmid capture system (PCS). The PCS donor-S transferred a pUC19 origin of replication and an ampicillin resistance marker into CpBV genomic DNA by in vitro transposition. Through PCS system, we were able to clone 53 genomic clones ranging from 0.1 to 25.5 kb and further they were classified into 29 segments by their sizes and restriction endonuclease patterns. Among them, a complete nucleotide sequence of CpBV-S28 segment was determined and 10 putative genes were predicted from this segment. Interestingly, 9 of 10 putative ORFs had high level of similarities with catalytic domain of protein tyrosine phosphatase. Also, ORF2807 showed similarity with EP1-like proteins of C. congregata polydnavirus.
Choi
Jae Young
JY
School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Republic of Korea.
Roh
Jong Yul
JY
Kang
Joong Nam
JN
Shim
Hee Jin
HJ
Woo
Soo Dong
SD
Jin
Byung Rae
BR
Li
Ming Shun
MS
Je
Yeon Ho
YH
D002874:Chromosome Mapping / Q000379:methods*; D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D016679:Genome, Viral; D010957:Plasmids / Q000235:genetics*; D018071:Polydnaviridae / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D004251:DNA Transposable Elements