人淋巴细胞免疫反应性生长激素基因调控序列的克隆与鉴定

王　辉　邓洁英　郑德先　史轶蘩

　　中国图书分类号　R392.11
　　摘　要　目的：克隆和鉴定人淋巴细胞免疫反应性生长激素(irGH)基因调控序列的核苷酸顺序。方法：通过PCR扩增出人淋巴细胞irGH基因5′近端的调控序列，然后将其与质粒PGEM7Zf(+)连接，经转化蓝白斑筛选、酶切鉴定，获得PGEM7Zf(+)-irGH DNA (-484-+2 bp)重组质粒，进行序列测定。结果：显示irGH基因5′近端的调控序列与垂体细胞GH基因的5′近端调控序列的核苷酸顺序完全相同。结论：该结果进一步证明免疫系统与神经内分泌系统的密切关系。
　　关键词　生长激素基因　调控序列　人淋巴细胞

Cloning and nucleotide sequencing of the 5′-flanking region of immunoreactive growth hormone gene of human lymphocyte

WANG Hui,DENG Jie-Ying,ZHENG De-Xian et al.
Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100730

　　　Abstract　Objective：Cloning and nucleotide sequencing of the 5′-flanking region of immunoreactive growth hormone gene of human 1ymphocyte.Methods:The immunoreactive growth hormone(irGH) DNA fragments were obtained by PCR with normal human peripheral blood lymphocyte DNA as template.Then,the plasmids of PGEM7Zf(+)-irGH DNA(-484-+2 bp) were constructed by ligation of irGH DNA with PGEM7Zf(+),transformation,blue/white color selection and restriction enzyme analysis.The 5′-flanking region of irGH were sequenced by using sanger s dideoxynucleitide chain-temination reaction.Results:It was demonstrated that the sequence data of 5′-flanking region(-484-+2 bp) in irGH gene was identical to that of pituitary GH gene.Conclusion:The results indicate a close relationship between immune system and neuroencrine system.
　　Key words　Growth hormone gene　Promoter　Lymphocytes

　　已发现经典的内分泌腺体、神经元、神经胶质细胞能够产生多种细胞因子，中枢及外周免疫器官也能产生多种激素和神经肽，此外还证实免疫系统和神经内分泌系统的细胞能表达细胞因子、激素及神经肽的受体［1］。淋巴细胞能分泌免疫反应性生长激素(Immunoreactive GH，irGH)，它是淋巴细胞的一种自分泌和旁分泌生长因子，在抗原性、分子量、生物活性及mRNA大小等方面与垂体GH相似。本实验室证实人淋巴细胞表达的irGH cDNA编码序列与垂体hGH cDNA的序列是相同的［2，3］，但目前有研究表明淋巴细胞分泌GH与垂体分泌GH的调控机制是不同的。本实验室为了从分子水平上探讨淋巴细胞irGH基因表达的调控机制，首先克隆irGH基因的5′上游调控序列，进行序列分析和鉴定。

1　材料与方法
1.1　菌株及质粒　DH5α及PGEM7Zf(+)质粒为本室保存。
1.2　试剂　内切酶(SmaI

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