Crawford R J, Wells J R
Mol Biol Rep. 1976 Nov;3(2):167-73. doi: 10.1007/BF00423231.
DNA, complementary to chicken globin mRNA was synthesized using either Avian Myeloblastosis virus reverse transcriptase, or E. coli DNA polymerase I. Transcriptase cDNA sediments at 9 S on sucrose gradients, and is 620 nucleotides in length, representing a complete copy of globin mRNA template. In contrast, Polymerase I cDNA sediments at 4 S, is 100 to 200 nucleotides in length, and is a copy of a small region at the 3'(poly A) end of globin mRNA. Similarly, Transcriptase cDNA and Polymerase I cDNA hybridize to globin mRNA template with characteristic, individual Crot1/2 values. The Crot1/2 value for Transcriptase cDNA hybridization is 7 X 10(-4) mol s 1(-1), and that for Polymerase I cDNA is 5 X 10(-3). This work shows that Avian Myeloblastosis virus reverse transcriptase can use Polymerase I cDNA to prime further cDNA synthesis along the mRNA template. The product of extended cDNA synthesis is identical in length and hybridization properties to oligo (dT) primed transcriptase cDNA.
利用禽成髓细胞瘤病毒逆转录酶或大肠杆菌DNA聚合酶I合成了与鸡珠蛋白mRNA互补的DNA。逆转录酶cDNA在蔗糖梯度上沉降系数为9S,长度为620个核苷酸,代表珠蛋白mRNA模板的完整拷贝。相比之下,聚合酶I cDNA沉降系数为4S,长度为100至200个核苷酸,是珠蛋白mRNA 3'(聚A)末端一个小区域的拷贝。同样,逆转录酶cDNA和聚合酶I cDNA以各自独特的Crot1/2值与珠蛋白mRNA模板杂交。逆转录酶cDNA杂交的Crot1/2值为7×10^(-4) mol s^(-1),聚合酶I cDNA的Crot1/2值为5×10^(-3)。这项工作表明禽成髓细胞瘤病毒逆转录酶可以利用聚合酶I cDNA沿mRNA模板引发进一步的cDNA合成。延伸的cDNA合成产物在长度和杂交特性上与寡聚(dT)引发的逆转录酶cDNA相同。