Abstract

cDNA for the VP4-coding RNA segment 4 of human rotavirus isolated from Korean patients Was synthesized and cloned (HRV-k41), and its nucleotide sequence was determined. Comparative analysis of the nucleotide sequence of JRV-k41 showed 90.6%, 86.6%, 74.6%, 66%, 70.1% and 65.4% homology to P1A[8](Wa), P1B[4](RV5), P2[6](1076), P3[9](AU1), P4[10](69M), and P[14](PA169) genotypes respectively. The deduced amino acid sequence homology of HRV-k41 to P1A[8](Wa), P1B[4](RV5), P2[6](1076), P3[9](AU1), P4[10](69M), and P[14](PA169) genotypes respectively. The deduced amino acid sequence homology of HRV-k41 to P1A[8](Wa), P1B[4](RV5), P2[6](1076), P3[9](AU1), P4[10](69M) and P[14](169) was 92.9% 89.7%. 75.8%, 64.1%, 70.6% and 64.3% respectively. These results suggest that HRV-k41 is closely related to the P1A genotype. Two trypsin cleavage sites (arginine 240 and arginine 246) and four cysteine residues (215, 317, 379, and 773) conserved in VP4 of all rotavirus strains were also found in JRV-k41. Similar to other virulent human rotaviruses, an additional trypsin cleavage site(lysine 245) was also detected in this strain. The cDNA of the VP4-coding RNA segment was cloned into pGEX-4T-3, an Escherichia coli expression vector, and it's expression was confirmed by Western-blot analysis.

• Keywords: cDNA; expression; human rotavirus; sequence; VP4