Seilhamer J J, Randall T L, Johnson L K, Heinzmann C, Klisak I, Sparkes R S, Lusis A J
California Biotechnology Inc., Mountain View 94043.
J Cell Biochem. 1989 Mar;39(3):327-37. doi: 10.1002/jcb.240390312.
We described previously the cloning and DNA sequence of the human gene encoding pancreatic phospholipase A2 [DNA 5, 519]. When pancreatic phospholipase A2 (PLA2) cDNA was used to screen a human genomic library, two classes of clones were obtained. One class encoded the pancreatic enzyme, and a second class encoded one exon of an apparently related PLA2. No additional PLA2 gene exons displayed sufficient homology to be detected by the probe. A homologous sequence in both rat and porcine genomic DNA was detected by DNA blot hybridization, and the corresponding gene fragments were cloned and sequenced. Within the deduced amino acid sequences, the presence of known functional residues along with the high degree of interspecies conservation suggests the genes encode a functional PLA2 enzyme form. The encoded sequence lacks Cys11, as do the "type II" viperid venom and other nonpancreatic mammalian PLA2 enzymes. The sequence is distinct from porcine intestinal PLA2 and appears not to be a direct homolog of the recently published rabbit ascites and rat platelet enzymes. Hybridization of DNA probes containing sequences from these genes to genomic DNA blots of mouse/human somatic cell hybrids permitted chromosomal assignment for both. The pancreatic gene mapped to human chromosome 12, and the homologous gene mapped to chromosome 1.
我们之前描述了编码人胰腺磷脂酶A2的基因的克隆及DNA序列[DNA 5, 519]。当用胰腺磷脂酶A2(PLA2)cDNA筛选人基因组文库时,获得了两类克隆。一类编码胰腺酶,另一类编码一个明显相关的PLA2的一个外显子。没有其他PLA2基因外显子显示出足够的同源性以被探针检测到。通过DNA印迹杂交在大鼠和猪的基因组DNA中检测到同源序列,并对相应的基因片段进行了克隆和测序。在推导的氨基酸序列中,已知功能残基的存在以及高度的种间保守性表明这些基因编码一种功能性PLA2酶形式。编码序列缺乏Cys11,“II型”蝰蛇毒和其他非胰腺哺乳动物PLA2酶也是如此。该序列与猪肠PLA2不同,似乎不是最近发表的兔腹水和大鼠血小板酶的直接同源物。含有这些基因序列的DNA探针与小鼠/人类体细胞杂种的基因组DNA印迹杂交,使得两者都能进行染色体定位。胰腺基因定位于人第12号染色体,同源基因定位于第1号染色体。