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目的:检测HBV亚基因型B和C的体外重组中间体。方法:将基因型B和C序列插入载体P lenti6/V5-D-topo-X后,共转染HepG2细胞,克隆,测定转染后HBV的核酸序列,而后用软件包RDP3Beta40进行序列比对。结果:发现存在3种重组中间体,重组中间体的重组位置为1 740-1 838至2 443-2 485。R1重组中间体的重组位置为nt2 170-2 172(CAC变成TGT)和nt2 188-2 189(GA变成AC);R2重组中间体的重组位置为nt1 740(A变成T),nt1 753(C变成T)和nt1 838(G变成A);R3重组中间体的重组位置为nt2 443(C变成T),nt2 425(A变成G),nt2 480(T变成C)和nt2 483(C变成T)。结论:HBV基因型B和C共感染可以产生重组,且重组位置在基因型B和C的前核心区和核心区。
Objective: To detect in vitro recombinant intermediates of HBV subgenotypes B and C. METHODS: HepG2 cells were cotransfected with genotypes B and C into P lenti6 / V5-D-topo-X vector, cloned and sequenced. The sequence of the HBV DNA was analyzed by software RDP3Beta40. Results: Three recombinant intermediates were found and the recombination intermediates were located at 1 740-1 838 to 2 443-2 485. The recombination sites of R1 recombination intermediates were nt2 170-2 172 (CAC changed to TGT) and nt2 188-2 189 (GA changed to AC). The recombination positions of R2 recombinants were nt1 740 (A changed to T), nt1 753 (C becomes T) and ntl 838 (G becomes A); the recombination positions of R3 recombination intermediates are nt2 443 (C becomes T), nt2 425 (A becomes G), nt2 480 (T becomes C ) And nt2 483 (C becomes T). CONCLUSION: Co-infection of HBV genotypes B and C can result in recombination with the recombination sites in the pre-core and core regions of genotypes B and C.