棉花恢复系中含有26S rRNA序列的GH18Rorf392基因克隆

Cloning of the Gene GH18Rorf392 Containing 26S rRNA Sequence from Cotton Restorer line

以Y18R×P30A杂交F1代自交材料播种后根据雄蕊形态学判断其育性,然后分别提取可育和不育材料现蕾3 ～5 d的幼蕾总RNA。通过抑制差减杂交的方法获得来自棉花恢复系的EST序列之后,以该EST序列设计引物再做5'和3'RACE,从而获得其全长cDNA序列。由于该基因来源于陆地棉(Gossypium hirsutum)恢复系Y18R,并且编码392个氨基酸的蛋白质,因此,将其命名为GH18Rorf392。该基因的5'端是一个全新的序列,3'端含有26S rRNA序列。该基因可能对研究棉花育性相关功能有帮助。

Y18R×P30A hybrid F1 generation then inbred, so judge the fertility according to stamen morphological character. Total RNA were extracted from young buds of 3-5 days from fertile and sterile cotton, respectively. EST sequence from cotton restorer line was obtained through suppression subtractive hybridization. Designing primers according to the EST sequence, then doing rapid amplification eDNA ends, so full length cDNA sequence was obtained. The gene was named by GH18Rorf392 because it came from Gossypium hirsutum restorer line Y18R and codes 392 amino acids. The 5＇ end of GH18Rorf392 is a new sequence and its 3＇ end contains 26S rRNA partial sequence. This gene can be used to study cotton fertility associated function.