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基因特异性引发互补DNA合成。

Gene specific priming of complementary DNA synthesis.

作者信息

Crawford R J, Wells J R

出版信息

Mol Biol Rep. 1976 Nov;3(2):167-73. doi: 10.1007/BF00423231.

Abstract

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.

摘要

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。

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