中南大学学报（医学版）
中南大學學報（醫學版）
중남대학학보（의학판）
JOURNAL OF CENTRAL SOUTH UNIVERSITY (MEDICAL SCIENCES)
2011年
2期
101-108
,共8页
陈俊意%黄爱龙%徐莉%陈典全%余虹%朱照静%黄祖春%杨宗发%陈立书%谭涛
陳俊意%黃愛龍%徐莉%陳典全%餘虹%硃照靜%黃祖春%楊宗髮%陳立書%譚濤
진준의%황애룡%서리%진전전%여홍%주조정%황조춘%양종발%진립서%담도
HBV%重组中间体%基因型B%基因型C%体外
HBV%重組中間體%基因型B%基因型C%體外
HBV%중조중간체%기인형B%기인형C%체외
hepatitis B virus%recombinant intermediate%genotype B%genotype C%in vitro
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Plenti6/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1740-1838至2443-2485.R1重组中间体的重组位置为nt2170-2172(CAC变成TGT)和 nt2188-2189(GA变成AC);R2重组中间体的重组位置为nt1740(A变成 T), nt1753(C变成 T )和 nt1838 (G 变成 A);R3重组中间体的重组位置为nt2443(C 变成 T), nt2425 (A 变成 G), nt2480( T变成 C)和nt2483 (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.
目的:檢測HBV亞基因型B和C的體外重組中間體.方法:將基因型B和C序列插入載體Plenti6/V5-D-topo-X後,共轉染HepG2細胞,剋隆,測定轉染後HBV的覈痠序列,而後用軟件包RDP3Beta40進行序列比對.結果:髮現存在3種重組中間體,重組中間體的重組位置為1740-1838至2443-2485.R1重組中間體的重組位置為nt2170-2172(CAC變成TGT)和 nt2188-2189(GA變成AC);R2重組中間體的重組位置為nt1740(A變成 T), nt1753(C變成 T )和 nt1838 (G 變成 A);R3重組中間體的重組位置為nt2443(C 變成 T), nt2425 (A 變成 G), nt2480( T變成 C)和nt2483 (C 變成T).結論:HBV基因型B和C共感染可以產生重組,且重組位置在基因型B和C的前覈心區和覈心區.
목적:검측HBV아기인형B화C적체외중조중간체.방법:장기인형B화C서렬삽입재체Plenti6/V5-D-topo-X후,공전염HepG2세포,극륭,측정전염후HBV적핵산서렬,이후용연건포RDP3Beta40진행서렬비대.결과:발현존재3충중조중간체,중조중간체적중조위치위1740-1838지2443-2485.R1중조중간체적중조위치위nt2170-2172(CAC변성TGT)화 nt2188-2189(GA변성AC);R2중조중간체적중조위치위nt1740(A변성 T), nt1753(C변성 T )화 nt1838 (G 변성 A);R3중조중간체적중조위치위nt2443(C 변성 T), nt2425 (A 변성 G), nt2480( T변성 C)화nt2483 (C 변성T).결론:HBV기인형B화C공감염가이산생중조,차중조위치재기인형B화C적전핵심구화핵심구.
Objective To detect the recombinant intermediates of hepatitis B virus (HBV) between genotype B and C in vitro. Methods Vector Plenti6/V5-D-topo-X was genetically modified by genotype B and C to transfect HepG2 cells. Then the HepG2 cells were amplified and sequence of the nucleic acid after the transinfection was tested and compared with RDP3Beta40 software package and bootscanning procedure in SimPlot program package. Results Three recombinant intermediates of HBV between genotype B and C were identified in vitro. Genotype C in the precore region plus the core gene spanning nucleotide positions from 1740-1838 to 2443-2485 contributed to the recombination with genotype B. Isolate R1 recombinant intermediate had 2 break points at nt2170-2172 and nt2188-2189. Nucleic acid changed from CAC to TGT and from GA to AC, respectively. Isolate R2 recombinant intermediate had a break point at nt1740-1 838, and 3 bases changed in different nucleic acid sites: from A to T at nt1740, from C to T at nt1753, and from G to A at nt1838, respectively. Isolate R3 recombinant intermediate had a break point at nt2443-2483, and 4 bases changed in different nucleic acid sites: from C to T at nt2443, from A to G at nt2452, from T to C at nt2480, and from C to T at nt2483, respectively. Conclusion The recombinant intermediates of HBV between genotype B and C have been detected in vitro and the changes have been identified in the precore region plus the core gene in genotype B and C.
