Sarih L, Araya A, Litvak S
Institut de Biochimie Cellulaire et Neurochimie du CNRS, Bordeaux, France.
FEBS Lett. 1988 Mar 28;230(1-2):61-6. doi: 10.1016/0014-5793(88)80642-2.
Bovine tRNA(Trp) can be partially hybridized to the avian myeloblastosis virus (AMV) 35 S RNA at 37 degrees C, in the presence of AMV RNA-dependent DNA polymerase (reverse transcriptase). This template-primer complex is active in the synthesis of viral cDNA. The size of the cDNA products synthesized in the in vitro reconstituted AMV system was determined by urea-polyacrylamide gel electrophoresis using a tRNA labelled at the 3'-end by yeast tRNA nucleotidyl transferase. The synthesized cDNA has a size of about 100 nucleotides and was shown by Southern blotting to be complementary to a specific sequence of the 5'-end of the retroviral genome. These results indicate that reverse transcriptase is able to anneal the exogenous primer tRNA at the 'primer-binding site' near the 5'-end of the long terminal repeat (LTR) of AMV RNA.
在禽成髓细胞瘤病毒（AMV）RNA依赖性DNA聚合酶（逆转录酶）存在的情况下，牛色氨酸转运RNA（tRNATrp）在37摄氏度时可与禽成髓细胞瘤病毒35S RNA部分杂交。这种模板-引物复合物在病毒互补DNA（cDNA）的合成中具有活性。在体外重建的AMV系统中合成的cDNA产物的大小，通过使用经酵母tRNA核苷酸转移酶在3'端标记的tRNA进行尿素-聚丙烯酰胺凝胶电泳来确定。合成的cDNA大小约为100个核苷酸，通过Southern印迹法显示其与逆转录病毒基因组5'端的特定序列互补。这些结果表明，逆转录酶能够使外源性引物tRNA在AMV RNA长末端重复序列（LTR）5'端附近的“引物结合位点”退火。
