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一些在组织和细胞分化中起重要作用的蛋白质包含锌指结构域 .为了克隆分离和研究与造血细胞分化和发育成熟相关的蛋白基因 ,利用编码C2H2型锌指蛋白结构域中部分保守氨基酸序列设计简并引物 ,以骨髓cDNA为模板 ,进行PCR扩增 ,得到若干新的锌指蛋白基因EST .用其中一条为探针筛选人骨髓cDNA文库 ,获得了一个新的锌指蛋白基因全长cDNA ,GenBank收录号为AF2 4 6 12 6 ,长 3888bp ,包括一个完整阅读框 ,编码 6 86个氨基酸 ,包括 17个典型的和 2个非典型的C2H2模体 ,命名为HZF2 .RNA印迹、人多组织mRNA斑点杂交分析结果显示 ,其在T淋巴细胞发育和定居的器官组织胸腺、淋巴结中有较高表达 ,在脾脏、胎肝有中度表达 ,在B淋巴细胞发育的骨髓中表达很低 ,在外周血几个淋巴细胞系中仅有极微量的表达 ,提示HZF2可能对于T淋巴细胞发育和增殖有重要功能 .该基因也在脑组织的若干部位、胎盘及肾上腺有较高表达 ,在多种其他组织细胞有微量表达 ,说明其可能对维持这些组织细胞的生理功能也起一定作用 .将编码HZF2读框的DNA顺序克隆到pEGFP N1载体中 ,转染 3T3细胞 ,证明表达的HZF2 GFP融合蛋白定位于细胞核 ,这与根据HZF2蛋白结构推测其可能作为DNA结合蛋白行使调节基因转录的功能是一致的 .
Some proteins that play an important role in tissue and cell differentiation include zinc finger domains.In order to isolate and study the protein genes involved in hematopoietic cell differentiation and developmental maturation, a partial conserved amino acid sequence encoding the C2H2-type zinc finger protein domain was designed Degenerate primers and bone marrow cDNA as a template for PCR amplification of a number of new zinc finger protein gene EST.With one of the probes as a screening of human bone marrow cDNA library obtained a full-length zinc finger protein gene cDNA, GenBank accession number AF2 4 6 12 6, length 3888bp, including a complete reading frame encoding 6 86 amino acids, including 17 typical and 2 atypical C2H2 motifs, named HZF2. RNA blotting, human multi-tissue The results of dot blot hybridization showed that it was highly expressed in the thymus and lymph nodes of T lymphocytes and colonized organ tissues, moderately expressed in spleen and fetal liver, and low in bone marrow of B lymphocytes. Only a very small amount of the expression of several lymphocytes in peripheral blood suggests that HZF2 may play an important role in the development and proliferation of T lymphocytes. Site, placenta and adrenal gland expression in a variety of other tissue cells trace expression, indicating that it may maintain the physiological function of these cells also play a role in. HZF2 encoding DNA sequence will be cloned into pEGFPN1 vector , Transfected 3T3 cells and proved that the expressed HZF2 GFP fusion protein localized in the nucleus, which is consistent with the function of HZF2 protein speculation that it may function as a DNA binding protein to regulate gene transcription.