Zakut R, Cohen J, Givol D
Nucleic Acids Res. 1980 Aug 25;8(16):3591-601. doi: 10.1093/nar/8.16.3591.
Poly(A)-containing mRNA from mouse myeloma MPC11 was transcribed into cDNA which was cloned in the PstI site of the plasmid pBR322. The transformants were screened by hybridization with a cDNA fragment, derived from plasmid p gamma(11)7, corresponding to the 5' portion of the constant region of MPC11 heavy chain. Several positive transformants were found to contain various lengths of the variable region of the heavy chain. We describe the structure and sequence of one of these clones, pV(11)2, which contains cDNA corresponding to the entire variable region of MPC11 heavy chain and extends to codon 248 in the constant region. The protein sequence deduced from the DNA sequence indicates that the variable region of MPC11 heavy chain contains 121 amino acids and belongs to subgroup II of mouse heavy chains. Comparison of this sequence with other heavy chain sequences suggests a J (joining) segment of 16 residues which overlaps five residues of the third hypervariable region. The cDNA sequence shows that there is no discontinuity between the end of the variable region and the beginning of the constant region.
从小鼠骨髓瘤MPC11中提取的含聚腺苷酸(Poly(A))的mRNA被转录成cDNA,该cDNA被克隆到质粒pBR322的PstI位点。通过与一个cDNA片段杂交筛选转化体,该cDNA片段源自质粒pγ(11)7,对应于MPC11重链恒定区的5'部分。发现几个阳性转化体含有不同长度的重链可变区。我们描述了其中一个克隆pV(11)2的结构和序列,它包含对应于MPC11重链整个可变区的cDNA,并延伸至恒定区的第248密码子。从DNA序列推导的蛋白质序列表明,MPC11重链的可变区包含121个氨基酸,属于小鼠重链的II亚组。将该序列与其他重链序列进行比较,发现一个由16个残基组成的连接(J)段,它与第三个高变区的五个残基重叠。cDNA序列表明,可变区末端与恒定区起始之间没有间断。