Lomonossoff G P, Shanks M, Matthes H D, Singh M, Gait M J
Nucleic Acids Res. 1982 Aug 25;10(16):4861-72. doi: 10.1093/nar/10.16.4861.
We have made use of the known sequence of the 5' ends of both CPMV RNAs to synthesise an oligodeoxynucleotide which can prime second-strand DNA synthesis on full-length cDNA copies of both RNAs. By priming synthesis in the presence of dideoxynucleoside triphosphates, we have determined the positions of the first AUG codons in each RNA. These occur at positions 115 and 207 on M and B RNA respectively. By using a cloned double-stranded DNA fragment derived from near the 5' end of M RNA as a primer additional sequence from the 5' terminal region of M RNA has been obtained.
我们利用已知的两种CPMV RNA 5'端序列合成了一种寡聚脱氧核苷酸，该寡聚脱氧核苷酸可引发这两种RNA全长cDNA拷贝上的第二链DNA合成。通过在双脱氧核苷三磷酸存在下引发合成，我们确定了每种RNA中第一个AUG密码子的位置。它们分别出现在M RNA和B RNA的第115位和第207位。通过使用源自M RNA 5'端附近的克隆双链DNA片段作为引物，获得了M RNA 5'末端区域的额外序列。
