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用5’RACE/3’RACE从海岛棉(Gossypium barbadense)开花后10 d的纤维细胞中分离获得了GbKTNl cDNA序列,全长2006 bp,包括113 bp的5’非翻译区、1563 bp的可读框及327 bp的3’非翻译区(不包括终止密码子TAA)。可读框编码521个氨基酸,蛋白质分子量约为55 kD,靠近GbKTNl C末端的233～414氨基酸残基间包含一个ATP结合功能区。Southern杂交证明,GbKTNl在海岛棉基因组中以单拷贝形式存在。半定量RT-PCR结合Southern杂交分析表明,GbKTNl在海岛棉根、茎、叶及纤维细胞中均有表达,以纤维细胞中的表达最高,叶中最低。利用裂殖酵母系统,初步分析了GbKTNl对细胞伸长的功能,发现该基因在酵母细胞中诱导表达后使细胞显著伸长,平均为诱导前的2.18倍,部分细胞呈不规则形状,这是首次在活体(in vivo)中证明KTN1与细胞的伸长有关。
The GbKTN1 cDNA sequence was isolated from fibroblasts of Gossypium barbadense by flow cytometry with 5’RACE / 3’RACE. The full-length cDNA sequence of GbKTN1 was 2006 bp, including 113 bp 5 ’untranslated region and 1563 bp Box and a 327 bp 3 ’untranslated region (excluding the stop codon TAA). The open reading frame encodes 521 amino acids with a molecular weight of about 55 kD. The 233 to 414 amino acid residues near the C terminus of GbKTN1 contain an ATP binding domain. Southern blotting demonstrated that GbKTN1 exists as a single copy in the island cotton genome. Semi-quantitative RT-PCR combined with Southern hybridization analysis showed that GbKTN1 was expressed in the roots, stems, leaves and fibroblasts of G. barbadense. The expression of GbKTN1 was highest in fibroblasts and lowest in leaves. The function of GbKTN1 on cell elongation was analyzed preliminarily by using the fission yeast system. It was found that the expression of GbKTN1 in yeast cells significantly prolonged, with an average of 2.18 times before induction and some cells were irregular, which is For the first time in vivo it was demonstrated that KTN1 is associated with cell elongation.