植物抗体基因工程：Ⅳ．马铃薯Y病毒小鼠中和抗体重链基因的序列分析及其植物表达  

Antibody Genetic Engineering in Plant: N. Sequencing of a Heavy Chain Gene against Potato Virus Y and Construction of Its Plant Expression Vector

作　　者：刘德虎[1] 陈三风[1] 李刚强 

机构地区：[1]中国农业科学院生物技术研究中心分子生物学室,北京农业大学生物学院微生物学系

出　　处：《高技术通讯》1996年第4期8-11,共4页Chinese High Technology Letters

基　　金：国家自然科学基金;世界青年科学基金

摘　　要：序列分析表明，马铃薯Ｙ病毒小鼠中和抗体重链基因（Ｇ９）全长为１５０５个核苷酸碱基（不包括Ｐｏｌｙ（Ａ）尾巴），其中包括５＇端非编码区１６个核苷酸碱基、编码重链信号肽的５７个核苷酸碱基、编码成熟蛋白的１３２９个核苷酸碱基和３＇端非编码区的１０３个核苷酸碱基。与已知的小鼠抗体Ｍｏｐｃ２１的重链基因（γ１）相比，核苷酸的序列同源性为９４．４％，由核苷酸序列所推导出的氨基酸序列同源性为９２．６％，其中可变区内（ＶＨ）的氨基酸序列同源性达７４．５％，这表明二者起源于同一种系基因。将完整的抗体重链基因正向插入植物表达载体ｐＢⅠ１２１中的原ＧＵＳ基因位置上，置于花椰莱花叶病毒３５Ｓ启动子控制之下，获得了能够在植物细胞内表达马铃薯Ｙ病毒中和抗体重链基因的植物表达载体ｐＹＨ。Sequence analysis showed that the immunoglobulin messenger RNA corresponding to G9 is 1505 nucleotides in length excluding the poly (A ) region, among which 16 bases cede for the 51 non-coding region, 57 for the leader sequence of the protein, 1329 for the mature protein and 103 for the 3' noncoding region. Comparison of G9 with the heavy chain gene of Mopc21 showed that they share a 94. 4 % identity in nucleotide sequence and 92. 6% identity in amino acid sequence. The heavy chain encoded by G9 was considered to be specific to PVY since only one type of heavy chain was expressed in the hybridoma. A binary vector, pYH, containing the light chain gene has been constructed for its expression in higher plants by introduction of the gene coding sequence into pBI121 instead of GUS gene.

关 键 词：马铃薯Y病毒 单克隆抗体 植物表达载体 

分 类 号：S532[农业科学—作物学]