植物学报
植物學報
식물학보
ACTA BOTANICA SINICA
2004年
11期
1301-1306
,共6页
吴传芳%安洁%何小佳%邓洁%洪志霞%刘超%吕鸿周%李宜瑾%王陈继%陈放%鲍锦库
吳傳芳%安潔%何小佳%鄧潔%洪誌霞%劉超%呂鴻週%李宜瑾%王陳繼%陳放%鮑錦庫
오전방%안길%하소가%산길%홍지하%류초%려홍주%리의근%왕진계%진방%포금고
朱顶兰凝集素(AVA)%单子叶甘露糖结合凝集素%简并引物%3′和5′RACE%同源克隆
硃頂蘭凝集素(AVA)%單子葉甘露糖結閤凝集素%簡併引物%3′和5′RACE%同源剋隆
주정란응집소(AVA)%단자협감로당결합응집소%간병인물%3′화5′RACE%동원극륭
Amaryllis vittata agglutinin (AVA)%monocot mannose-binding lectin%degenerate primers%3′and 5′RACE%homology alignment
运用同源克隆的方法设计简并引物,通过3′和5′RACE技术,从石蒜科植物朱顶兰(Amaryllis vittata Ait)总RNA中克隆了编码此凝集素(AVA)的全长cDNA序列.该基因全长686 bp,起始密码子位于第41～43 bp,终止密码子位于515～517 bp处,开放阅读框长474 bp,编码158个氨基酸,包含信号肽序列、成熟蛋白序列和C-末端剪切序列的前体蛋白.成熟蛋白由109个氨基酸残基组成,分子量为11.9kD.成熟蛋白在氨基酸水平上与雪花莲凝集素、水仙凝集素、石蒜凝集素和君子兰凝集素分别有73.4%、85.3%、80.7%和83.5%的同源性;朱顶兰凝集素的分子模式显示其与雪花莲凝集素有极其相似的三维结构;在Blocks数据库中检索AVA蛋白氨基酸序列的结构域,发现有3个凝集素功能结构域,并具有3个典型的甘露糖专一结合位点盒(QDNY).
運用同源剋隆的方法設計簡併引物,通過3′和5′RACE技術,從石蒜科植物硃頂蘭(Amaryllis vittata Ait)總RNA中剋隆瞭編碼此凝集素(AVA)的全長cDNA序列.該基因全長686 bp,起始密碼子位于第41～43 bp,終止密碼子位于515～517 bp處,開放閱讀框長474 bp,編碼158箇氨基痠,包含信號肽序列、成熟蛋白序列和C-末耑剪切序列的前體蛋白.成熟蛋白由109箇氨基痠殘基組成,分子量為11.9kD.成熟蛋白在氨基痠水平上與雪花蓮凝集素、水仙凝集素、石蒜凝集素和君子蘭凝集素分彆有73.4%、85.3%、80.7%和83.5%的同源性;硃頂蘭凝集素的分子模式顯示其與雪花蓮凝集素有極其相似的三維結構;在Blocks數據庫中檢索AVA蛋白氨基痠序列的結構域,髮現有3箇凝集素功能結構域,併具有3箇典型的甘露糖專一結閤位點盒(QDNY).
운용동원극륭적방법설계간병인물,통과3′화5′RACE기술,종석산과식물주정란(Amaryllis vittata Ait)총RNA중극륭료편마차응집소(AVA)적전장cDNA서렬.해기인전장686 bp,기시밀마자위우제41～43 bp,종지밀마자위우515～517 bp처,개방열독광장474 bp,편마158개안기산,포함신호태서렬、성숙단백서렬화C-말단전절서렬적전체단백.성숙단백유109개안기산잔기조성,분자량위11.9kD.성숙단백재안기산수평상여설화련응집소、수선응집소、석산응집소화군자란응집소분별유73.4%、85.3%、80.7%화83.5%적동원성;주정란응집소적분자모식현시기여설화련응집소유겁기상사적삼유결구;재Blocks수거고중검색AVA단백안기산서렬적결구역,발현유3개응집소공능결구역,병구유3개전형적감로당전일결합위점합(QDNY).
A new mannose-binding agglutinin gene was cloned from bulbs of Amaryllis vittata Ait. The fulllength cDNA of A. vittata agglutinin (AVA) was 686 bp. The start codon of ava cDNA was at 41-43 bp and the stop codon was at 515-517 bp. Analysis in the BLAST of GenBank showed that ava gene encodes a protein precursor composed of a signal peptide, mature protein and C-terminal amino acid cleavage sequence. The mature protein of AVA includes 109 amino acid residues and the molecular weight is 11.9 kD. The homologous analysis showed that the identity between AVA and Galanthus nivalis agglutinin, Narcissus hybrid cultivar agglutinin, Lycoris radiata agglutinin, Clivia miniata agglutinin are 73.4%, 85.3%, 80.7%, 83.5%,respectively. Molecular modeling of AVA indicated that its three-dimensional structure strongly resembles that of the snowdrop agglutinin. Blocks' analysis revealed that the deduced amino acid sequence of AVA has three functional domains specific for agglutination and three carbohydrate-binding boxes (QDNY).
