Cloning of a ribonucleotide reductase gene of the herpes simplex virus type 2 strain G.
The ribonucleotide reductase (RR) 2 gene of the HSV-2 strain G was cloned, sequenced, and expressed in an E. coli cell. The RR2 gene was located on the PstI 2.4 kb fragment, which was cloned and sequenced. The ORF of the gene was 1,011 bp and its termination codon was TAG; also, the CATATAA sequence was present in the promoter of the RR2 gene. A Poly A signal sequence (AATAAA) was found in the 3'-noncoding region. The RR2 proteins that were produced in the E. coli and Vero cells were confirmed using a Western blot analysis. SDS-PAGE revealed that the molecular weights of the fusion-RR2 that was produced in the E. coli cells were approximately 24 kDa and 38 kDa in the Vero cells. The RR2 proteins were soluble. The differences in the molecular weights might be due to modifications in the Vero cells.
Kim
Hee Jin
HJ
Department of Biological Sciences, Konkuk University, Seoul 143-701, Korea.
Lee
Si Kyung
SK
Byun
Si Myung
SM
Lee
Hyung Hoan
HH
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D018258:Herpesvirus 2, Human / Q000235:genetics*; D008969:Molecular Sequence Data; D012264:Ribonucleotide Reductases / Q000235:genetics*
D012264:Ribonucleotide Reductases