柠条锦鸡儿CkLEA4基因克隆及表达分析

Clone and Expression Analysis of CkLEA4 fromCaragana korshinskii Kom.

LEA蛋白是一种与植物抗逆相关的胚胎发育晚期丰富蛋白。该研究从已构建的柠条锦鸡儿干旱胁迫抑制削减杂交文库中筛选到1条LEA蛋白编码基因的部分序列,用RACE技术扩增得到该基因cDNA全长并对其进行了克隆。测序表明该基因cDNA长870bp,其中开放阅读框长510bp,编码169个氨基酸,推测蛋白分子量为17.03kD,等电点为9.3,是一种亲水蛋白。序列比对和系统进化分析表明,该基因属于LEA4基因家族成员,命名为CkLEA4。实时荧光定量PCR检测发现,CkLEA4基因在干旱、ABA和NaCl处理下均受到不同程度的诱导,说明CkLEA4基因可能与柠条锦鸡儿响应逆境胁迫有关。

Late embryogenesis abundant （LEA） proteins could be induced in plants under stress conditions. In this study,an LEA encoding gene fragment was isolated from the suppression subtractive hybridization library of Caragana korshinskii Kom. ,and the full length cDNA was cloned by rapid amplification of cD- NA ends technique. Sequencing results showed that the full length cDNA was 870 bp,with an open reading frame of 510 bp. It encodes a protein composed of 169 amino acids with a calculated molecular mass of 17.03 kD,and a theoretical pI of 9.3. The deduced protein is strongly hydrophilic. Sequence and phyloge- netic analysis indicated that the protein belongs to the LEA4 subfamily,therefore it was designated as Ck- LEA4. Real-time quantitative PCR analysis showed that the transcript of CkLEA4 was induced strongly under different treatments, including ABA, salt and drought. These results indicated that CkLEA4 might be involved in stress responses of C. korshinskii Kom.