Tan K L, Webb G C, Baker R T, Board P G
Molecular Genetics Group, John Curtin School of Medical Research, Australian National University, Canberra.
Genomics. 1995 Jan 20;25(2):381-7. doi: 10.1016/0888-7543(95)80037-m.
Until recently the Theta-class glutathione S-transferases (GSTs) were largely overlooked due to their low activity with the model substrate 1-chloro-2,4-dinitrobenzene (CDNB) and their failure to bind to immobilized glutathione affinity matrices. Little is known about the number of genes in this class. Recently, Pemble et al. (Biochem J. 300: 271-276, 1994) reported the cDNA cloning of a human Theta-class GST, termed GSTT1. In this study, we describe the molecular cloning of a cDNA encoding a second human Theta-class GST (GSTT2) from a lambda gt11 human liver 5'-stretch cDNA library. The encoded protein contains 244 amino acids and has 78.3% sequence identity with the rat subunit 12 and only 55.0% identity with human GSTT1. GSTT2 has been mapped to chromosome 22 by somatic cell hybrid analysis. The precise position of the gene was localized to subband 22q11.2 by in situ hybridization. The absence of other regions of hybridization suggests that there are no closely related sequences (e.g., reverse transcribed pseudogenes) scattered throughout the genome and that if there are closely related genes, they must be clustered near GSTT2. Southern blot analysis of human DNA digested with BamHI shows that the size of the GSTT2 gene is relatively small, as the coding sequence falls within a 3.6-kb BamHI fragment.
直到最近,θ类谷胱甘肽S-转移酶(GSTs)在很大程度上被忽视了,因为它们对模型底物1-氯-2,4-二硝基苯(CDNB)的活性较低,并且无法与固定化谷胱甘肽亲和基质结合。关于这类基因的数量知之甚少。最近,Pemble等人(《生物化学杂志》。300:271 - 276, 1994)报道了一种人θ类GST(称为GSTT1)的cDNA克隆。在本研究中,我们描述了从λgt11人肝5'-延伸cDNA文库中克隆编码第二种人θ类GST(GSTT2)的cDNA。编码的蛋白质含有244个氨基酸,与大鼠亚基12的序列同一性为78.3%,与人类GSTT1的同一性仅为55.0%。通过体细胞杂交分析,GSTT2已被定位到22号染色体。通过原位杂交,该基因的确切位置被定位到22q11.2亚带。没有其他杂交区域表明在整个基因组中没有紧密相关的序列(例如,反转录假基因),并且如果存在紧密相关的基因,它们必须聚集在GSTT2附近。用BamHI消化的人DNA的Southern印迹分析表明,GSTT2基因的大小相对较小,因为编码序列位于一个3.6kb的BamHI片段内。