Bao Jinku, Wu Chuanfang, An Jie, Gao Shun, Zhao Xi, Chang Liqing, Rong Yanzhen, Wang Chenji, Chen Fang
College of Life Science, Sichuan University, Chengdu 610064, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2004 Oct;21(5):812-8.
The monocot mannose-binding lectin can inhibit HIV from infecting the target cells. The total RNA of Zephyranthes grandiflora was extracted and reversely transcribed into cDNA. Degenerate primers were designed based on the conserved regions of other monocot mannose-binding agglutinins by homology alignment. The 694bp full-length cDNA of Zephyranthes grandiflora agglutinin (ZGA) was cloned by RT-PCR, 3' and 5' RACE (rapid amplification of cDNA ends). The start codon and stop codon of ZGA were at 37-39bp and 529-531bp respectively. The NCBI Blast analysis result showed that ZGA gene encoded a protein precursor with signal peptide, mature protein and C-terminal cleavage sequence. The mature ZGA protein contained 106 amino acids residues and its molecular weight was 11.6KD. The percentages of identity of the deduced mature ZGA protein with those of Galanthus nivalis agglutinin, Narcissus hybrid cultivar agglutinin, Lycoris radiate agglutinin and Clivia miniata agglutinin were 71.8%, 81%, 81.8% and 84.5%, respectively. Blocks analysis revealed that ZGA had three functional domains and three mannose-binding boxes (QDNY).
单子叶甘露糖结合凝集素可抑制HIV感染靶细胞。提取了葱兰的总RNA并反转录成cDNA。通过同源比对,基于其他单子叶甘露糖结合凝集素的保守区域设计了简并引物。通过RT-PCR、3'和5' RACE(cDNA末端快速扩增)克隆了葱兰凝集素(ZGA)694bp的全长cDNA。ZGA的起始密码子和终止密码子分别位于37 - 39bp和529 - 531bp处。NCBI Blast分析结果表明,ZGA基因编码一个具有信号肽、成熟蛋白和C末端切割序列的蛋白质前体。成熟的ZGA蛋白含有106个氨基酸残基,分子量为11.6KD。推导的成熟ZGA蛋白与雪花莲凝集素、水仙杂交品种凝集素、石蒜凝集素和君子兰凝集素的同一性百分比分别为71.8%、81%、81.8%和84.5%。结构域分析显示ZGA有三个功能结构域和三个甘露糖结合框(QDNY)。