摘要
用带有鸡溶菌酶基因增强子（Ｅ）和启动子（Ｐ）的编码细菌氯霉素乙酰基转移酶（ＣＡＴ）的融合基因（ＥＰＣ），在其上游区、下游区或在上游和下游区同时插入鸡溶菌酶基因３＇端核基质附着区（ＭＡＲ）的不同表达载体ＭＥＰＣ、ＥＰＣＭ和ＭＥＰＣＭ，稳定转染鸡ＨＤ１１Ｐｒｏｍａｃｒｏｐｈａｇｅ同源细胞系，筛选不同表达载体稳定整合后的细胞株，检测ＣＡＴ表达水平，确定其表达基因拷贝数，从而分析鸡溶菌酶基因３＇端ＭＡＲ对基因表达的影响。稳定整合表达和瞬时表达实验结果表明，鸡溶菌酶基因３＇端ＭＡＲ在同源细胞系对基因表达没有激活作用，与该基因５＇端ＭＡＲ在同源或异源细胞系中能激活基因表达形成鲜明对照，说明５＇瑞ＭＡＲ和３＇端ＭＡＲ在调节鸡溶菌酶基因表达上存在一种协调关系。
Matrix attachment regions(MARs) have been identified in severalgenes.Nuclear MARs in genomic DNA are thought to be involved in nearly all impor-tant processes of the nucleus,for instance,the organization of chromatin loop-domains,DNA replication, DNA repairing; RNA transcription and processing.The MARs of thechicken lysozyme gene were identified at the boundaries of the 'active' chromatin do-main.The MAR element located 5'of the chicken lysozyme gene has been shown to me-diate elevated,position-less dependent expression of genes which stably transfected intochicken or heterologous cells.In this paper,chicken HD11 cells were stably transfectedeither with a construct (EPC)containing the chicken lysozyme gene enhancer(E)andpromoter(P)fused to the reporter gene(C)encoding bacterial chloramphenical acetyltransferase( CAT)ene or with the constructs( MEPCM, MEPC, EPCM)in whichEPC transcription units were flanked by chicken lysozyme gene3'MAR.Results showedthat the 3' MAR from the chicken lysozyme gene couldn't activate the expression oftransfected genes in homologous cells.
出处
《中国兽医学报》
CAS
CSCD
1996年第3期212-217,共6页
Chinese Journal of Veterinary Science
基金
中德农业合作研究资助项目
关键词
MAR
同源细胞系
基因表达
溶菌酶
鸡
MAR
homologous cells
gene expression
lysozyme
chicken Institute for Small AnimalResearch,FederalAgricultural Research Center,Doenbergstr.25-27,29223 Celle,Germany
