IN VITRO MUTAGENESIS OF CHICKEN HISTONE & HMG GENES

HMG-14和HMG-17是普遍存在的非组蛋白染色体蛋白 与核小体核心颗粒结合的物质。他们组成了一个家庭 已被报道与活性染色质结合的染色体蛋白质。 为了了解这些蛋白质在体内的功能，我们扰乱了 HMG-14a和HMG-17在禽类B细胞系DT40中均有表达。我们的结果 表明基因敲除细胞的表型没有明显变化。 相对于亲本DT40细胞。此外，没有补偿 观察到细胞内hMG-14b或组蛋白水平的变化。 缺少HMG-14a和HMG-17。On-D凝胶电泳法 分析发现，一种主要的表达蛋白在 基因敲除细胞。凝胶和蛋白质分析方法的发展 直接在转移膜上被用来鉴定这一点 特定的地点。

HMG-14 and HMG-17 are ubiquitous non-histone chromosomal proteins that bind to nucleosome core particles. They form a family of chromosomal proteins that have been reported to bind active chromatin. To understand the in vivo function of these proteins, we disrupted both HMG-14a and HMG-17 in an avian B-cell line DT40. Our results indicate that the knockout cells show no obvious change in phenotype with respect to the parental DT40 cells. Furthermore, no compensatory changes in HMG-14b or histone protein levels were observed in cells lacking both HMG-14a and HMG-17. Upon 2-D gel electrophoretic analysis, a major expresssed protein was found to be activated in the knockout cells. Developing methods for analyzing proteins in gels and directly on transfer membranes are being employed to identify this particular spot.