Strätling W H, Dölle A, Sippel A E
Biochemistry. 1986 Jan 28;25(2):495-502. doi: 10.1021/bi00350a033.
The chromatin structure encompassing the lysozyme gene domain in hen oviduct nuclei was studied by measuring the partitioning of coding and flanking sequences during chromatin fractionation and by analyzing the nucleosome repeat in response to micrococcal nuclease digestion. Following micrococcal nuclease digestion, nuclei were sedimented to obtain a chromatin fraction released during digestion (S1) and then lysed in tris(hydroxymethyl)aminomethane-(ethylenedinitrilo)tetraacetic acid-[ethylenebis(oxyethylenenitrilo)]tetraacetic acid and centrifuged again to yield a second solubilized chromatin fraction (S2) and a pelleted fraction (P2). By dot-blot hybridization with 14 specific probes, it is found that the fractionation procedure defines three classes of sequences within the lysozyme gene domain. The coding sequences, which partition with fraction P2, are flanked by class I flanking sequences, which partition with fractions S1 and P2 and which extend over 11 kilobases (kb) on the 5'side and probably over about 4 kb on the 3' side. The partitioning of class II flanking sequences, which are located distal of class I flanking sequences, is different from that of class I flanking sequences. Coding sequences lack a canonical nucleosome repeat, class I flanking sequences possess a disturbed nucleosome repeat, and class II flanking sequences generate an extended nucleosomal ladder. Coding and class I flanking sequences are more readily digested by micrococcal nuclease than class II flanking sequences and the inactive beta A-globin gene. In hen liver, where the lysozyme gene is inactive, coding and class I flanking sequences fractionate into fractions S2 and P2.(ABSTRACT TRUNCATED AT 250 WORDS)
通过在染色质分级分离过程中测量编码序列和侧翼序列的分配情况，以及分析微球菌核酸酶消化后的核小体重复序列，对鸡输卵管细胞核中包含溶菌酶基因结构域的染色质结构进行了研究。微球菌核酸酶消化后，将细胞核沉淀以获得消化过程中释放的染色质部分（S1），然后在三（羟甲基）氨基甲烷 - （乙二胺四乙酸） - [亚乙基双（氧乙烯基腈）]四乙酸中裂解并再次离心，得到第二个可溶染色质部分（S2）和沉淀部分（P2）。通过与14种特异性探针的斑点杂交发现，分级分离程序在溶菌酶基因结构域内定义了三类序列。与沉淀部分P2一起分配的编码序列，其侧翼是I类侧翼序列，I类侧翼序列与部分S1和P2一起分配，在5'侧延伸超过11千碱基（kb），在3'侧可能延伸约4 kb。位于I类侧翼序列远端的II类侧翼序列的分配与I类侧翼序列不同。编码序列缺乏典型的核小体重复序列，I类侧翼序列具有受干扰的核小体重复序列，II类侧翼序列产生延伸的核小体梯带。与II类侧翼序列和无活性的βA - 珠蛋白基因相比，编码序列和I类侧翼序列更容易被微球菌核酸酶消化。在溶菌酶基因无活性的鸡肝脏中，编码序列和I类侧翼序列分级分离到部分S2和P2中。（摘要截短于250字）
