| [Objective]:Different HBV genotypes may have different biological characteristics, viral genotype may be associated with long-term progression of infection and reactions to interferon therapy.Currently full-length sequences of HBV of China is very few, This study was to establish a rapid and efficient method for receiving HBV genome cloning, determining viral gene sequence and getting a number of full-length sequences od HBV.then we take the sequence analysis Microsoft to do type analysis with the resulting sequences, analyzes the differences and association of different genotype.accordingly, the HBV gene pool can be ruched,and can provide reference basis for HBV variation HBV drugs and vaccines research.[Methods] To ollected100Infected serum samples of chronic positive HBV infecter,extract HBV full genome,design appropriate specific primers by reference of the100HBV full-length sequence from Genbank,amplified the extractors, and receiving HBV PCR production, linked them with appropriate carriers,picked out the right monoclonal with Blue-white screening,sequenced them and analyzed with micro-softs[Resμlt]:established a new method of attachment differentiated with traditional way for TA cloning which could Improve the efficiency and success rate of connection,Directly connected the full-length HBV sequence and T vector. Among the51full-length sequences submited to NCBI, there are18genotype B(35.29%) and33genotype C (64.70%),no genotype B+C.[Conclusion]:Connection with efficient cloning method that can greatly simplify the steps of connection of fragment with vector and improve the efficiency and success ratio.they were all genotype B and genotype C by analyze the cloning51HBV full-sequence. Because of the small sample size and population mobility, it was unable to analyze the epidemiological characteristics of geographical areas. |
