Cloning And Expression Of Neuraminidase Gene Of Influenza Virus H1N1

Almost every year,Influenza can cause different scales epidemic in different countries,which leaded to severe damage to human health and economic development.Neuraminidase(NA) is an important structure protein of Influenza Virus,and a key target which can induce humoral immunity. So far,NA inhibitor has become a main drug to cure Influenza.Vaccines based on NA protein and NA gene are also in developing state.Owing to Influenza vaccine based on NA is a promising field,so the objective of this study is to get correct recombinant PET21-NA1 whose NA1 gene is from A/WSN/1933(H1N1) using gene cloning technique and purified interest protein.The competent protein will be injected into mouse which can produce specific immune response,and monoclonal antibody technique will be used to get monoclone,and antibody will be purified and checked by ELISA.Another goal is to inject correct recombinant into mouse through intramuscular injection or particle gun,and the relative protein will be expressed in vivo which can induce specific immune response,and finally nucleic acid vaccine of NA gene will be gained.In this study,DNA was obtained by PCR using cDNA of NA1 from H1N1 as template.Clone vector PMD18-T ligated with PeR product was transformed into E.coli DH5α,and positive clone strain was identified.Then, expression vector PET21a-d(+) ligated with NAI gene was transformed into E.coli BL21,and positive recombinant was selected and identified by enzymes and PCR.Ni-NTA agarose was used to purify interest protein.This study will help development of vaccine based on Neuraminidase,and facilitate understanding to H1N1 Influenza virus.