Yasuda K, Kondoh H, Okada T S, Nakajima N, Shimura Y
Nucleic Acids Res. 1982 May 11;10(9):2879-91. doi: 10.1093/nar/10.9.2879.
Double-stranded DNA was synthesized from delta-crystallin mRNA prepared from lens fibers of 15-day-old chick embryos and cloned at the Pst I site of the plasmid pBR322. Using the cloned cDNA and single-stranded cDNA as hybridization probes, a number of genomic DNA fragments containing delta-crystallin gene sequences have been cloned from the partial and complete EcoRI digests of chick brain DNA. One of the clones from the partial digests contains a DNA fragment that consists of four EcoRI fragments of 7.6 kb, 4.0 kb, 2.6 kb, and 0.8 kb. The gene sequences reside in the (5')7.6 kb - 0.8 kb - 4.0 kb (3') fragments. Electron microscopy has provided evidence that the cloned DNA fragment includes the entire gene sequences complementary to delta-crystallin mRNA except for the 3' terminal poly(A) tail, and that the delta-crystallin gene is interrupted by at least 13 intervening sequences. Another clone contains a genomic fragment that consists of two EcoRI fragments of 3.0 kb and 11 kb. The DNA fragment in the latter clone represents a different delta-crystallin gene, as judged by restriction endonuclease mapping and by electron microscopy.
双链DNA是由从15日龄鸡胚晶状体纤维制备的δ-晶体蛋白mRNA合成的,并克隆到质粒pBR322的Pst I位点。使用克隆的cDNA和单链cDNA作为杂交探针,从鸡脑DNA的部分和完全EcoRI消化物中克隆了许多含有δ-晶体蛋白基因序列的基因组DNA片段。部分消化物中的一个克隆包含一个DNA片段,该片段由7.6 kb、4.0 kb、2.6 kb和0.8 kb的四个EcoRI片段组成。基因序列位于(5')7.6 kb - 0.8 kb - 4.0 kb (3')片段中。电子显微镜提供了证据,表明克隆的DNA片段包含除3'末端聚(A)尾外与δ-晶体蛋白mRNA互补的整个基因序列,并且δ-晶体蛋白基因被至少13个间隔序列中断。另一个克隆包含一个基因组片段,该片段由3.0 kb和11 kb的两个EcoRI片段组成。通过限制性内切酶图谱分析和电子显微镜判断,后一个克隆中的DNA片段代表一个不同的δ-晶体蛋白基因。