包含鸡组蛋白基因的基因组克隆的分离

Harvey R P, Wells J R

Nucleic Acids Res. 1979 Dec 11;7(7):1787-98. doi: 10.1093/nar/7.7.1787.

We have used enriched chicken histone cDNA to select genomal clones from a chicken library. Because the cDNA probe also contained other sequences, a further screening of positive plagues with negative probes eliminated most non-histone gene clones. One 'positively-selected' genomal clone, lambda CH-01, hybridised with cloned sea-urchin histone genes and also detected histone genes in EcoRI-digested genomal sea-urchin DNA. Limited DNA sequencing of HaeIII fragments identified two sequences within the coding region of chicken histone H2A. A third fragment predicted an amino acid sequence with strong homology to an H1 histone sequence.

我们利用富集的鸡组蛋白cDNA从鸡文库中筛选基因组克隆。由于cDNA探针还包含其他序列，用阴性探针进一步筛选阳性噬菌斑可去除大多数非组蛋白基因克隆。一个“正向选择”的基因组克隆λCH-01，与克隆的海胆组蛋白基因杂交，并且在经EcoRI酶切的海胆基因组DNA中也检测到了组蛋白基因。对HaeIII片段进行有限的DNA测序，确定了鸡组蛋白H2A编码区内的两个序列。第三个片段预测的氨基酸序列与H1组蛋白序列有很强的同源性。