Bonifer C, Vidal M, Grosveld F, Sippel A E
Zentrum für Molekulare Biologie Universität Heidelberg, FRG.
EMBO J. 1990 Sep;9(9):2843-8. doi: 10.1002/j.1460-2075.1990.tb07473.x.
A 21.5 kb DNA fragment carrying the entire chicken lysozyme gene locus was introduced into the germ line of mice. The fragment contains the transcribed region plus 11.5 kb 5'-flanking and 5.5 kb 3'-flanking sequences including all known cis-regulatory elements and the 5' and 3' attachment elements (A-elements) which define the borders of the DNase I sensitive chromatin domain. All sequences which adopt a DNase I hypersensitive chromatin conformation in vivo are present on the construct. Seven founder mice were analysed. All of these expressed chicken lysozyme RNA at high levels specifically in macrophages, as is the case in the donor species. Expression levels are dependent on the copy number of integrated genes indicating that a complete gene locus, as defined by its chromatin structure, functions as an independent regulatory unit when introduced into a heterologous genome.
一个携带完整鸡溶菌酶基因位点的21.5 kb DNA片段被导入小鼠生殖系。该片段包含转录区域以及11.5 kb的5'侧翼和5.5 kb的3'侧翼序列，包括所有已知的顺式调控元件以及定义DNase I敏感染色质结构域边界的5'和3'附着元件（A元件）。构建体上存在所有在体内呈现DNase I超敏感染色质构象的序列。对7只奠基小鼠进行了分析。所有这些小鼠都在巨噬细胞中特异性地高水平表达鸡溶菌酶RNA，就像在供体物种中一样。表达水平取决于整合基因的拷贝数，这表明由其染色质结构定义的完整基因位点在被导入异源基因组时作为一个独立的调控单元发挥作用。
