Erickson P F, Maxwell I H, Su L J, Baumann M, Glode L M
Department of Medicine, University of Colorado Health Sciences Center, Denver 80262.
Biochem J. 1990 Jul 15;269(2):335-40. doi: 10.1042/bj2690335.
A cDNA clone for cystathionine gamma-lyase was isolated from a rat cDNA library in lambda gt11 by screening with a monospecific antiserum. The identity of this clone, containing 600 bp proximal to the 3'-end of the gene, was confirmed by positive hybridization selection. Northern-blot hybridization showed the expected higher abundance of the corresponding mRNA in liver than in brain. Two further cDNA clones from a plasmid pcD library were isolated by colony hybridization with the first clone and were found to contain inserts of 1600 and 1850 bp. One of these was confirmed as encoding cystathionine gamma-lyase by hybridization with two independent pools of oligodeoxynucleotides corresponding to partial amino acid sequence information for cystathionine gamma-lyase. The other clone (estimated to represent all but 8% of the 5'-end of the mRNA) was sequenced and its deduced amino acid sequence showed similarity to those of the Escherichia coli enzymes cystathionine beta-lyase and cystathionine gamma-synthase throughout its length, especially to that of the latter.
通过用单特异性抗血清筛选，从λgt11载体中的大鼠cDNA文库中分离出胱硫醚γ-裂解酶的cDNA克隆。通过阳性杂交选择确认了该克隆的身份，它包含基因3'端近端600bp的片段。Northern印迹杂交显示，相应的mRNA在肝脏中的丰度高于大脑。通过与第一个克隆进行菌落杂交，从质粒pcD文库中又分离出另外两个cDNA克隆，发现它们的插入片段分别为1600bp和1850bp。其中一个通过与对应于胱硫醚γ-裂解酶部分氨基酸序列信息的两个独立寡脱氧核苷酸池杂交，被确认为编码胱硫醚γ-裂解酶。对另一个克隆（估计代表mRNA 5'端除8%以外的所有部分）进行了测序，其推导的氨基酸序列在全长上与大肠杆菌酶胱硫醚β-裂解酶和胱硫醚γ-合酶的氨基酸序列相似，尤其与后者相似。
