Mathews A, Llewellyn D J, Wu Y, Dennis E S
CSIRO Division of Plant Industry, Canberra City, Australia.
Plant Mol Biol. 1996 Mar;30(5):1035-9. doi: 10.1007/BF00020813.
A full-length cDNA encoding the 206 amino acid open reading frame of a trypsin/chymotrypsin inhibitor abundant in the corms of giant taro (Alocasia macrorrhiza) was isolated. An internal fragment was cloned using degenerate primers corresponding to a region of the mature protein sequence and the 'rapid amplification of cDNA ends' (RACE) method used to generate a composite cDNA sequence. The length of the cDNA was close to the predicted size of the corresponding transcript deduced from northern blot analysis of corm mRNA. The inhibitor was expressed strongly in the mature corm, at low levels in leaf blades and petioles but not in roots. Southern blot analysis of the giant taro DNA indicated that this inhibitor is encoded by a small multigene family and this was further supported by the isolation of two different sequence classes from corm cDNA using primers to the ends of the composite sequence.
分离出了一个全长cDNA，其编码一种在巨型芋头（海芋）球茎中大量存在的胰蛋白酶/糜蛋白酶抑制剂的206个氨基酸的开放阅读框。使用与成熟蛋白序列区域对应的简并引物克隆了一个内部片段，并采用“cDNA末端快速扩增”（RACE）方法生成了一个复合cDNA序列。该cDNA的长度接近从球茎mRNA的Northern印迹分析推断出的相应转录本的预测大小。该抑制剂在成熟球茎中强烈表达，在叶片和叶柄中表达水平较低，但在根中不表达。对巨型芋头DNA的Southern印迹分析表明，这种抑制剂由一个小的多基因家族编码，从球茎cDNA中使用复合序列末端的引物分离出两种不同的序列类别，进一步支持了这一点。
