DETECTION OF COMPLEMENT PROTEIN MESSENGER-RNA IN HUMAN ASTROCYTES BY THE POLYMERASE CHAIN-REACTION
DETECTION OF COMPLEMENT PROTEIN MESSENGER-RNA IN HUMAN ASTROCYTES BY THE POLYMERASE CHAIN-REACTION
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DOI：
10.1016/0165-0270(92)90076-p
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发表时间：
1992-12-01
影响因子：
3
通讯作者：
SADLON, TA
中科院分区：
文献类型：
--
作者：
GORDON, DL;AVERY, VM;SADLON, TA
设计针对补体蛋白C3、B因子和I因子的特异性引物，通过聚合酶链式反应从培养的人脑星形胶质细胞中扩增出c DNA。扩增产物大小适中，分别为506bp、885bp和146bp，并用改进的化学发光检测系统进行Southern印迹分析。检测灵敏度高，约6250个细胞的DNA扩增产物清晰可见。C3和B因子以前曾被报道是由小鼠星形胶质细胞产生的；然而，这是第一次有报告表明人类星形胶质细胞合成了C3、B因子和I因子。这些结果表明，聚合酶链式反应是检测人脑星形胶质细胞补体替代途径蛋白的一种简单而灵敏的方法。
Primer sets specific for complement proteins, C3, factor B and factor I, were designed and used to amplify cDNA from cultured human astrocyte mRNA by the polymerase chain reaction. Appropriately sized PCR products of 506 bp, 885 bp and 146 bp, respectively, were generated and specificity was confirmed with Southern blotting using an enhanced chemiluminescence detection system. The sensitivity of detection was high, with amplified product from cDNA of approximately 6250 cells readily visualized. C3 and factor B have previously been reported to be produced by murine astrocytes; however, this is the first report indicating synthesis of C3, factor B and factor I by human astrocytes. These results indicate that PCR is a simple and sensitive technique to detect mRNA transcripts for proteins of the alternative pathway of complement in human astrocytes.