EXPRESSION AND FUNCTION OF PORCINE CLASS I MHC GENES IN TRANSGENIC MICE

猪I类MHC基因在转基因小鼠中的表达和功能

• 批准号: 3813471
• 负责人: D SINGER
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3813471
• 关键词: DNA; chromosome deletion; gene expression; genetic enhancer element; genetic manipulation; genetically modified animals; histocompatibility; histocompatibility antigens; interferons; laboratory mouse; major histocompatibility complex; swine; transfection; transplantation

Porcine class I major histocompatibility (MHC) genes have been introduced into the genome of C57BL/10 mice in order to study their in vivo patterns of expression and to analyze the ability of their products to function. One transgenic line which contains a classical class I MHC gene (PD1) expresses SLA antigen on its cell surface skin grafts from the B1O.PD1 transgenic are rejected by normal C57BL/10 mice, indicating that the foreign SLA antigen is recognized as a functional transplantation antigen Expression of PD1 parallels that observed in the swine, indicating that its expression is regulated. In vivo treatment of the transgenic with sigma/beta-interferon increases PD1 expression in a number of tissues. To further characterize regulatory DNA sequence elements responsible for tissue specific expression of class I gene, a series of 5' deletion mutants of PD1 was generated. DNA constructs containing the deletions mutants ligated to a reporter gene, CAT, were introduced into transgenic animals and assessed for their patterns of expression. Within 1.1 kb upstream of transcriptional initiation, negative and positive regulatory elements were identified, some of which function in a tissue specific fashion. In particular, a complex element consisting of an overlapping enhancer and silencer demonstrates marked tissue specificity. Transgenic lines have been generated containing a swine class I gene PD7, which is closely related to PD1. The two lines differ in the extent of non-coding 5' flanking sequence which has been introduced with the PD7 gene. Analysis of PD7 expression in the two lines has revealed the presence of tissue specific silencer associated with the gene. The difference in extent of 5' flanking sequences in the two constructs results not only in differential levels of antigen expression but different rates of graft rejection.

猪I类主要组织相容性（MHC）基因已被引入 进入C57 BL/10小鼠的基因组以研究其体内模式 并分析其产品的功能。一 含有经典I类MHC基因（PD 1）的转基因系表达 从B1O.PD1转基因皮肤移植物的细胞表面SLA抗原是 正常C57BL/10小鼠排斥，表明外源SLA抗原 被认为是功能性移植抗原PD1的表达 与在猪中观察到的相似，表明其表达是 监管.用σ/β-干扰素体内治疗转基因 增加PD1在许多组织中的表达。为了进一步表征 负责组织特异性表达的调节DNA序列元件 通过对Ⅰ类基因5 ′端缺失突变体的筛选，获得了一系列5 ′端缺失突变体。DNA 含有与报道基因连接的缺失突变体的构建体， CAT被引入转基因动物中，并评估其 表达模式。转录上游1.1 kb内 启动，负和正调控元件被确定，一些 其以组织特异性方式发挥功能。特别是，一个复杂的 由重叠的增强子和沉默子组成的元件表明， 明显的组织特异性。 已经产生了含有猪I类基因PD7的转基因系， 它与PD1密切相关。这两条线的区别在于 已与PD7一起引入的非编码5 '侧翼序列 基因对两个细胞系中PD7表达的分析揭示了存在PD7表达。 与该基因相关的组织特异性沉默者。的差异 两种构建体中5 ′侧翼序列的延伸不仅导致 抗原表达水平不同，但移植率不同 排斥反应