美洲商陆一种抗真菌蛋白的cDNA克隆及序列分析  

cDNA Cloning and Sequencing of an Antifungal Proteinfrom the Seeds of Phytolacca americana

作　　者：鄢波[1,2] 马志刚[1,2] 黄兴奇 王铃仙[1,2] 胡忠 

机构地区：[1]云南省农科院生物技术研究所 [2]中国科学院昆明植物研究所

出　　处：《云南植物研究》1998年第3期276-278,共3页Acta Botanica Yunnanica

基　　金：云南省自然科学基金

摘　　要：根据美洲商陆（Ｐｈｙｔｏｌａｃａａｍｅｒｉｃａｎａ）种子中一种抗真菌蛋白ＰＡＦＰ（Ｈｕｅｔａｌ，１９９１）的Ｎ端部分氨基酸顺序，设计、合成一条５’端寡核苷酸引物，通过３’－ＲＡＣＥ技术从种子总ＲＮＡ扩增出一约３５０ｂｐ的ｃＤＮＡ片段，成功地克隆到ｐＧＥＭ－Ｔ载体系统中。序列分析表明该ｃＤＮＡ含有１１４ｂｐ的编码区，由此推导的３８个氨基酸序列有３６个与ＰＡＦＰ的序列相同，仅在第３５、３６位氨基酸分别由Ｃｙｓ、Ｌｙｓ替代了后者的Ｇｌｎ和Ｉｌｅ。１１４ｂｐ的编码序列也被克隆。该ｃＤＮＡ可能编码一种新的３８个氨基酸的抗真菌蛋白。Basing on the amino acid sequences of 7 kD antifungal protein from Phytolacca americana ( Hu Zhong et al,1991 ), We designed a pair of degenerated primers. By using 3'-RACE technique, we have amplified a cDNA fragment about 350 bp from the total RNA of the seeds of Phytolacca americana. PCR product was cloned into pGEM-T vectoy system directly, and the cDNA sequences was then determinated. The results indicated that the cDNA fragment contains an encoding region about 114 bp, and that the deduced amino acid sequences is the same as the 36 amino acids of the PAFP protein,except the two amino acids of the No.35 and No.36, at which Gln and Ile is replaced by Cys and Lys respectively. In addition, the encoding region was cloned. The cDNA may encodea a new antifungal protein of 38 amino acids.

关 键 词：美洲商陆 抗真菌蛋白 基因克隆 

分 类 号：Q943[生物学—植物学]