Wall R, Gilmore-Hebert M, Higuchi R, Komaromy M, Paddock G, Strommer J, Salser W
Nucleic Acids Res. 1978 Sep;5(9):3113-28. doi: 10.1093/nar/5.9.3113.
Recombinant DNA clones have been generated from mouse myeloma MOPC 21 immunoglobulin kappa light chain mRNA. Complementary DNA (cDNA) synthesized on kappa light chain mRNA by reverse transcriptase was made double stranded and inserted into the bacterial plasmid vector, pMB9. Approximately 70 tetracycline-resistant transformed colonies containing kappa light chain mRNA sequences were identified by colony hybridization. Five of these recombinant clones were selected and characterized. Three clones contain both kappa light chain constant and variable region sequences. Two of these three recombinant clones have been shown to include all of the kappa light chain constant and variable region coding sequences. Another of the five selected recombinant clones contain kappa light chain constant region sequences. The remaining characterized clone appears to be derived from sequences at the 5'-end of kappa light chain mRNA, possibly extending to the terminal cap structure.
已从小鼠骨髓瘤MOPC 21免疫球蛋白κ轻链mRNA生成重组DNA克隆。通过逆转录酶在κ轻链mRNA上合成的互补DNA(cDNA)被双链化并插入细菌质粒载体pMB9中。通过菌落杂交鉴定出约70个含有κ轻链mRNA序列的四环素抗性转化菌落。从这些菌落中挑选并鉴定了5个重组克隆。其中3个克隆同时包含κ轻链恒定区和可变区序列。这3个重组克隆中的2个已被证明包含所有的κ轻链恒定区和可变区编码序列。5个挑选出的重组克隆中的另一个包含κ轻链恒定区序列。其余经鉴定的克隆似乎源自κ轻链mRNA 5'端的序列,可能延伸至末端帽结构。