Nilson J H, Thomason A R, Horowitz S, Sasavage N L, Blenis J, Albers R, Salser W, Rottman F M
Nucleic Acids Res. 1980 Apr 11;8(7):1561-73. doi: 10.1093/nar/8.7.1561.
Poly(A)-containing RNA from the bovine anterior pituitary has been used as a template for the enzymatic synthesis of double-stranded cDNA. The resulting double-stranded cDNA was inserted into the Pst I site of pBR322 with the oligo(dG)-oligo(dC) tailing technique and subsequently cloned in E. coli chi 1776. Clones containing sequences complementary to prolactin mRNA were identified by colony hybridization with partially purified prolactin cDNA. A 250 base pair sequence from one prolactin positive clone was extensively characterized and shown to contain the coding information for amino acids 119-192 of authentic bovine prolactin. The recombinant DNA from this clone was covalently attached to diazotized aminocellulose and used to purify prolactin mRNA from a mixture of mRNAs.
来自牛脑垂体前叶的含Poly(A)RNA已被用作酶促合成双链cDNA的模板。通过寡聚(dG)-寡聚(dC)加尾技术，将所得双链cDNA插入pBR322的Pst I位点，随后克隆到大肠杆菌chi 1776中。通过与部分纯化的催乳素cDNA进行菌落杂交，鉴定出含有与催乳素mRNA互补序列的克隆。对一个催乳素阳性克隆的250个碱基对序列进行了广泛表征，结果表明其包含真实牛催乳素第119 - 192位氨基酸的编码信息。来自该克隆的重组DNA与重氮化氨基纤维素共价连接，并用于从mRNA混合物中纯化催乳素mRNA。
