摘要： 为了研究CkF5H基因在柠条锦鸡儿中木质素生物合成途径方面中的生物学功能，以我国西北干旱荒漠地区重要的豆科饲用灌木柠条锦鸡儿为材料，克隆了1个木质素合成相关的CkF5H基因，该基因cDNA全长1 679 bp，gDNA全长3 495 bp，具有2个外显子，1个内含子，编码520个氨基酸，GenBank登录号HQ829862。通过染色体步移的技术克隆了1 550 bp的启动子序列，对启动子序列上的响应元件进行了分析。结果显示，具有昼夜节律响应元件、防御与胁迫响应元件、赤霉素顺式作用元件和热响应元件等。CkF5H富含二级结构，其中α螺旋占39.81%，β折叠占15.96%，不规则卷曲占44.23%，并预测其三维结构。对CkF5H基因推导的氨基酸序列预测分析得出，CkF5H蛋白等电点为6.45，分子量为58.09 ku，不稳定系数45.72，是不稳定蛋白，总平均亲水性-0.143，属亲水性蛋白。因此，克隆柠条锦鸡儿的CkF5H基因并详细分析，可为进一步研究该基因提供理论依据。

关键词: 柠条锦鸡儿, F5H, 基因克隆, 木质素, RACE

Abstract: CkF5H gene related to lignin synthesis was cloned from Caragana korshinskii Kom., an important leguminous forage shrub in the arid desert region of Northwest China. The full length of the gene cDNA was 1 679 bp. A total of 3 495 bp gDNA sequences of CkF5H gene of Caragana korshinskii were obtained,which had two exons,one intron,encoded 520 amino acids,GenBank accession number HQ829862. And the promoter sequence of 1 550 bp CkF5H gene was cloned by chromosome walking technique. The response elements on the promoter sequence were analyzed. The results showed that there were circadian rhythm response elements,defense and stress response elements,gibberellin cis-acting elements. CkF5H was rich in secondary structure,α-spiral accounted for 39.81%,β-fold accounted for 15.96%,irregular curl accounted for 44.23%,and predicted its three-dimensional structure.The predicted deduced amino acid sequence of CkF5H gene showed that the protein had an isoelectric point of 6.45,a molecular weight of 58.09 ku and an unstable coefficient of 45.72,which was an unstable protein with a total average hydrophilicity of -0.143 and a hydrophilic protein. Therefore,Cloning Caragana korshinskii Kom. CkF5H gene and detailed analysis will provide a theoretical basis for further study of the gene.

Key words: Caragana korshinskii Kom., F5H, Gene cloning, Lignin, RACE

中图分类号: 

• Q943.2