Cloning of an almost full-length chicken conalbumin double-stranded cDNA.
Cloning of an almost full-length chicken conalbumin double-stranded cDNA.
复制标题
克隆几乎全长的鸡伴清蛋白双链 cDNA。
作者：
M. Cochet;F. Perrin;F. Gannon;A. Krust;P. Chambon;G. McKnight;D. C. Lee;K. E. Mayo;R. Palmiter
从富含伴清蛋白mRNA（con-mRNA）的蛋鸡输卵管mRNA制备物合成鸡伴清蛋白双链cDNA（con-dscDNA）。通过平端连接将dscDNA插入质粒pBR 322的Sal I位点，该位点已用DNA聚合酶I修复，以在插入片段的每一侧产生Taq I位点。在细菌转化后，一个杂交重组体，pBR 322-con 1，它含有最大的插入dscDNA（约2350 bp），显示特异性杂交的RNA被翻译成伴清蛋白。电子显微镜检查con-mRNA和pBR 322-con 1 DNA之间的杂交分子表明，插入的con-dscDNA几乎是一个全长的双链转录的伴清蛋白mRNA。
Chicken conalbumin double-stranded cDNA (con-dscDNA) was synthesized from a laying hen oviduct mRNA preparation enriched for conalbumin mRNA (con-mRNA). The dscDNA was inserted by blunt-end ligation into the Sal I site of plasmid pBR322 which had been repaired with DNA polymerase I to create Taq I sites on each side of the inserted fragment. After bacterial transformation, one hybrid recombinant, pBR322-con1, which contains the largest inserted dscDNA (about 2350 bp) was shown to hybridize specifically to the RNA which is translated into conalbumin. Electron microscopic examination of hybrid molecules between con-mRNA and pBR322-con1 DNA indicate that the inserted con-dscDNA is an almost full-length double-stranded transcript of conalbumin mRNA.