Deev S M, Chuvpilo S A, Karlyshev A V, Mukhamedov R S, Polianovskiĭ O L
Genetika. 1982;18(6):888-95.
Restriction analysis of hybrid plasmids from a number of clones allowed to screen out the plasmid p8-1. A detailed analysis of this plasmid and that of the inserted DNA was made using HhaII, AluI, Sau96I, Sau3A, MspI and BspI restriction endonucleases. The results demonstrated that the inserted DNA corresponds to the 3'-nontranslated region and to a portion of the C fragment of the light-chain immunoglobulin gene. Establishing the partial structure of the recombinant plasmid exhibited a complete coincidence of the inserted DNA 3'-terminus and the primary structure of the light-chain mRNA synthesized by MOPC-21 myeloma cells. This sequence corresponds to the amino acid sequence in the light-chain constant region (207-214 residues). The analysis of p8-1 plasmid showed that nucleotides of pBR322 plasmid at positions from 376 to 618 were deleted. The deletion might be predetermined by the plasmid property to be amplified more readily than all the other hybrid plasmids which were not deleted.
对多个克隆的杂交质粒进行限制性分析,从而筛选出质粒p8-1。使用HhaII、AluI、Sau96I、Sau3A、MspI和BspI限制性内切酶对该质粒以及插入的DNA进行了详细分析。结果表明,插入的DNA对应于免疫球蛋白轻链基因的3'-非翻译区和C片段的一部分。重组质粒部分结构的确定显示,插入DNA的3'-末端与MOPC-21骨髓瘤细胞合成的轻链mRNA的一级结构完全一致。该序列对应于轻链恒定区的氨基酸序列(207 - 214个残基)。对p8-1质粒的分析表明,pBR322质粒位于376至618位的核苷酸被缺失。这种缺失可能由该质粒比所有其他未缺失的杂交质粒更易于扩增的特性所决定。