大肠杆菌DNA聚合酶I合成与乳蛋白mRNA互补的DNA
Synthesis of DNA complementary to the mRNAs for milk proteins by E. coli DNA polymerase I.
作者信息
Houdebine L M
出版信息
Nucleic Acids Res. 1976 Mar;3(3):615-30. doi: 10.1093/nar/3.3.615.
E.Coli DNA polymerase I (Klenow subfragment) was used for the synthesis of complementary DNA with the mRNAs for rabbit milk proteins as templates. The cDNA formed, contained 200 nucleotides and represented about 20% of the mRNA template. The cDNA was hybridized specifically to the mRNA templates. The Klenow subfragment of the E.Coli DNA polymerase I was as efficient as the avian myeloblastosis virus reverse transcriptase in the synthesis of cDNA. The mean size of the cDNA fragments obtained with the Klenow enzyme proved to be 70% of the value obtained with the AMV reverse transcriptase and at least twice the value generally obtained with the complete E.Coli DNA polymerase I. The cDNA was used for the detection and the quantification of the mRNA template in various RNA fractions.
大肠杆菌DNA聚合酶I（克列诺夫亚片段）用于以兔乳蛋白的mRNA为模板合成互补DNA。形成的cDNA含有200个核苷酸，约占mRNA模板的20%。该cDNA与mRNA模板特异性杂交。大肠杆菌DNA聚合酶I的克列诺夫亚片段在cDNA合成中与禽成髓细胞瘤病毒逆转录酶一样高效。用克列诺夫酶获得的cDNA片段的平均大小证明是用禽成髓细胞瘤病毒逆转录酶获得的值的70%，并且至少是通常用完整的大肠杆菌DNA聚合酶I获得的值的两倍。该cDNA用于检测和定量各种RNA组分中的mRNA模板。
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