Li Yuying, Zhang Zheng, Liang Aihua, Wang Zhuanhua
Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, People's Republic of China.
DNA Seq. 2006 Jun;17(3):203-7. doi: 10.1080/10425170600699760.
Based on the amino acid information of trypsin inhibitor of buckwheat (Fagopyrum Esculentum Moench), degenerated primers were designed and a full-length cDNA sequence named BTIomega1 (Buckwheat Trypsin Inhibitor) was amplified from the leaves RNA by using RT-PCR and rapid amplification of cDNA ends (RACE) methods. Sequence analysis shows that the 392 bp cDNA contained an open reading frame (ORF) of 216 bp, encoding 72 amino acids residues. The deduced amino acid sequence exhibits 96 and 93% homology with BWI-1 and BTI-2, a natural trypsin inhibitor from buckwheat seeds. Southern blotting suggested that three copies of BTIomega1 gene existed in the buckwheat genome. Moreover, a predicted secondary structure and 3D-structural model was constructed by homology modeling. To our knowledge, this is the first all-round report of the gene BTIomega1. The novel BTIomega1 gene has been submitted to the GeneBank under Accession No. DQ289792.
基于荞麦（苦荞麦）胰蛋白酶抑制剂的氨基酸信息，设计了简并引物，并采用逆转录聚合酶链反应（RT-PCR）和cDNA末端快速扩增（RACE）方法，从叶片RNA中扩增出一个名为BTIomega1（荞麦胰蛋白酶抑制剂）的全长cDNA序列。序列分析表明，392 bp的cDNA包含一个216 bp的开放阅读框（ORF），编码72个氨基酸残基。推导的氨基酸序列与来自荞麦种子的天然胰蛋白酶抑制剂BWI-1和BTI-2分别具有96%和93%的同源性。Southern杂交表明，荞麦基因组中存在3个BTIomega1基因拷贝。此外，通过同源建模构建了预测的二级结构和三维结构模型。据我们所知，这是关于BTIomega1基因的首次全面报道。该新的BTIomega1基因已提交至基因库，登录号为DQ289792。
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