Objective: To determine the nucleotide sequences of M and NS genes of A/Goose/Guangdong/2/96(H5N1) virus and also to compare them with the sequences of A/HK/156/97(H5N1) strain for revealing the relationship between the two viruses, as well as for setting up a solid base for studying M and NS genes of influenza A viruses in the future.
Methods: Virion RNA was transcribed into cDNA by reverse transcriptase, cDNA was amplified by PCR, the products of PCR were purified. Afterward, RNA sequence analysis was performed by the dideoxynucleotide chain termination method using synthetic oligodeoxynucleotide primers.
Results: The segment length of A/Goose/Guangdong/2/96(H5N1) virus RNA 7 is 1,027 nucleotides. It codes M1 (252 amino acids) and M2 (97 amino acids) proteins. However, the segment length of RNA 8 of A/Goose/Guangdong/2/96(H5N1) virus is 890 nucleotides coding NS1 (230 amino acids) and NS2 (121 amino acids) proteins. The amino acid sequence homologies of M1, M2, NS1 and NS2 protein molecules between A/Goose/Guangdong/2/9(H5N1) and A/HK/156/97 (H5N1) viruses are 97.6%, 92.8%, 65.7% and 76.9%, respectively.
Conclusion: The lengths of A/Goose/Guangdong/2/96(H5N1) virus RNA 7 and RNA 8 and 1,027 are 890 nucleotides, respectively. These two RNA segments belong to those of avian influenza A virus. The A/HK/156/97(H5N1) virus RNA 7 and RNA 8 segments are not derived from those of A/Goose/Guangdong/2/96(H5N1) virus directly.