Siddiqui A, Sattler F, Robinson W S
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4664-8. doi: 10.1073/pnas.76.9.4664.
DNA of hepatitis B virus (HBV) of hepatitis B surface antigen (HBsAg) subtype adw2 made fully double stranded by the virion DNA polymerase and radiolabeled either by the virion DNA polymerase reaction or by nick-translation with 32P-labeled deoxynucleoside triphosphates was used to establish a map of restriction endonuclease cleavage sites by the method double and triple enzyme digestion and to determine the relative positions of several unique physical features of this DNA. The five restriction sites for enzyme HincII, the two sites each for BamHI, Ava I, and Bgl II, and the single sites for EcoRI, Pst I, Hpa I, and Taq I were positioned relative to each other. Within this map, the single-stranded region in HBV DNA has been localized and the locations of nicks in each strand (a and b) have been determined with respect to restriction sites on the circular map. Comparison of restriction endonuclease cleavage patterns of DNAs of HBV of HBsAg subtypes adw2, ayw3, and adrq+ revealed consistent differences among subtypes and occasional differences within subtypes.
由病毒粒子DNA聚合酶使乙肝表面抗原(HBsAg)亚型adw2的乙肝病毒(HBV)DNA完全双链化,并通过病毒粒子DNA聚合酶反应或用32P标记的脱氧核苷三磷酸进行切口平移进行放射性标记,以此通过双酶切和三酶切方法建立限制性内切酶切割位点图谱,并确定该DNA几个独特物理特征的相对位置。确定了酶HincII的五个限制性位点、BamHI、Ava I和Bgl II各自的两个位点以及EcoRI、Pst I、Hpa I和Taq I的单个位点的相对位置。在该图谱中,已定位了HBV DNA中的单链区域,并已确定每条链(a和b)上切口相对于环状图谱上限制性位点的位置。对HBsAg亚型adw2、ayw3和adrq+的HBV DNA的限制性内切酶切割模式进行比较,结果显示各亚型之间存在一致差异,且各亚型内部偶尔也有差异。