• 作者： 吳妍華; 羅時成; 邱建興; 丁令白; 劉武哲; 朱廣邦
• 作者服務機構： 陽明醫學院生化研究所; 陽明醫學院微生物及免疫研究所; 陽明醫學院醫技系
• 中文摘要： 本篇論文中，我們將國內B型肝炎病患血液中鄧氏顆粒的B型肝炎病毒基因複殖于大腸菌 JM103。我們探用 pUC8
  為輸送分子。二個重組體命名為pTWLl及pTWL2被發現帶有3.2kbp ECORI 片段，並經Southern hybridization方法
  證實確實帶有B型肝炎病毒基因。
  我們亦將部份鄧氏顆粒中的B型肝炎病毒基因的單鏈部份以內生性DNA聚合醉修補缺口，再以BamHI限制?切割
  ，插入pUC8的BamHIr處。四個菌落pTWS1，pTWS2，pTWS3，pTWS4被證實帶有1.4kbp BamHI片段，此片段上有
  B型肝炎病毒表面抗原基因。但僅pTWSl質體上的表面抗原基因的轉錄方向和Lac起動子方向一致。
  本篇論文並將質體pTWSl及pTWLl上之B型肝炎病毒基因的限制?拼圖決定。實驗結果顯示出病毒基因上，限制?
  XbaI，HpaI的切割處僅有一處，而限制? SstI，SalI，SmaI，KpnI, HindIII?無切割點。限制?BamHI，HincII，
  BglII有多切割點，且切割點的排列位置和已發表的亞型adw及adw 類似。
• 英文摘要： The EcoRI fragment of hepatitis B virus (HBV) DNA isolated from human blood plasma Daneparticles were inserted into plasmid pUC8 EcoRI site and transformed into E. coli JM103 host. Two re-combinants pTWL1 and pTWL2 were found to carry 3.2 kbp fragment and proved to have HBV genome bySouthern hybridization method. The 1.4 kbp Bam HI fragment which carried the hepatitis B viral surface antigen (HBsAg) gene, ob-tained via BamHI digestion of Dane particles DNA which was made fully double stranded by endogenousDNA polymerase reaction, was also inserted into plasmid pUC8 BamHI site. Four recombinant clones,pTWSl，pTWS2, pTWS3, and pTWS4 were found. Only one of the clones pTWSl carried the HBsAg genein a correct orientation with respect to the lac promoter sequence. The physical mapping of HBV DNA was performed with several restriction endonucleases. Our re-sults indicated that the HBV DNA insert contains unique XbaI and HpaI cleavage sites and lacks the cleav-age sites for the HindIII, SmaI, KpnI, SalI, and SstI endonucleases. The locations of BamHI, BgIII, andHincII endonucleases cleavage sites within the cloned HBV DNA of the pTWLl plasmid were similar to thatHBV DNA of adw and adw subtypes.
• 中文關鍵字： hepatitis B virus (HBV); hepatitis B surface antigen (HBs Ag) restriction endonuclease; southern blot; subtype; cloning.
• 英文關鍵字： --