Cloning and nucleotide sequence analysis of the nucleocapsid protein gene of NDV strain V_4

Objective To study biochemical function and therapy of the NP(nucleocapsid protein) gene of NDV(new castle disease virus).Methods The purified genome RNA of NDV were used for template.According to the reported NP gene sequence of NDV strain Beaudettec C,a pair of 19 mer primers was designed and synthesized.Then the NP gene of NDV strain V 4 was amplified by reverse transcription polymerase chain reaction(RT PCR).The amplified products were analyzed by agarose gel electrophoresis,a specific fragment about 1.5 kb as expected appeared.The RT PCR product of strain V 4 was cloned into the pUC119,then the positive clone was tested by restriction endonuclease analysis and it was sequenced by Sanger's method.Results The result suggested that it was the NP gene of NDV strain V 4.Conclusion The NP gene sequence of NDV V 4 as compared with published NP gene sequence of NDV Beaudettec C,La Sota and D 26 ,affinal are 90.64%, 90.17 %,98.03%;difference rate of the sequence of amino acid of the NP gene are 4.50%,5.93%,2.45%.