CHARACTERIZATION AND EXPRESSION OF THE PROSTATIC ARGININE ESTERASE GENE, A CANINE GLANDULAR KALLIKREIN
CHARACTERIZATION AND EXPRESSION OF THE PROSTATIC ARGININE ESTERASE GENE, A CANINE GLANDULAR KALLIKREIN
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DOI：
10.1089/dna.1991.10.49
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发表时间：
1991-01-01
影响因子：
3.1
通讯作者：
DUBE, JY
中科院分区：
文献类型：
--
作者：
CHAPDELAINE, P;GAUTHIER, E;DUBE, JY
从用犬肝DNA构建的基因组文库中克隆了前列腺癌精氨酸酯酶基因。所选克隆含有约17kb的插入片段，包括精氨酸酯酶mRNA的完整编码部分(5个外显子和4个内含子)，在起始点上游2kb，在聚腺苷酸化位点下游12kb。内含子-外显子边界与所有已知的哺乳动物激肽释放酶基因相同。推导出的氨基酸序列与其他激肽释放酶有很高的同源性(51-61%)，不仅在前列腺中表达，而且在各种动物的胰腺中表达。5‘-侧翼序列含有潜在的调控元件，如变异型TATA盒(TTTAAA)、CCAAT盒、SP1转录因子结合位点(GGGCGG)和两个类似糖皮质激素反应元件的TGTCCT基序。用精氨酸酯酶基因不同部分的DNA探针对扩增出的487bp的cDNA5‘端片段进行Southern杂交分析表明，犬基因组中存在两到三个同源基因，而在先前的研究中，用400bp的精氨酸酯酶cDNA3’端片段检测到单一条带。这些结果表明精氨酸酯酶基因属于一个小激肽释放酶基因家族。精氨酸酯酶mRNA主要在前列腺中表达，但在其他几个组织中也有极低水平的表达(大约低1000倍)，包括肝脏、股薄肌、肾脏和胰腺。
The prostatic arginine esterase gene was isolated from a genomic library prepared with dog liver DNA in lambda-EMBL3. The selected clone contained an insert of approximately 17 kb which included the whole coding portion of arginine esterase mRNA (5 exons plus 4 introns), 2 kb upstream from the initiation site and 12 kb downstream from the polyadenylation site. The intron-exon boundaries were identical to all known mammalian kallikrein genes. The deduced amino acid sequence indicated a high degree of identity (51-61%) with other kallikreins expressed not only in the prostate but also in the pancreas of various animal species. The 5'-flanking sequences contained potential regulatory elements such as a variant TATA box (TTTAAA), a CCAAT box, a SP1 transcriptional factor binding site (GGGCGG), and two TGTCCT motifs resembling glucocorticoid response elements. Southern blot analysis with an amplified cDNA fragment of 487 bp corresponding to the 5' portion of the mRNA and with a DNA probe from a different portion of the arginine esterase gene indicated the presence of two to three homologous genes in the canine genome while in a previous study a single band was detected using a 400-bp arginine esterase cDNA corresponding to the 3' portion of the mRNA. These results suggest that the arginine esterase gene belongs to a small kallikrein gene family. Arginine esterase mRNA is expressed primarily in the prostate but also at an extremely low level (approximately a thousandfold less) in several other tissues including the liver, the gracilis thigh muscle, the kidney, and the pancreas.