论文部分内容阅读
利用RT-PCR和RACE技术,从新疆栽培扁桃‘鹰嘴’花药中获得了一个全长的SFB(S-hap-lotype-specific F-boxgene)基因(GenBank登录号为FJ362525),命名为AcSFBd。该基因全长1125bp,编码374个氨基酸,在N末端含有一个大约50个氨基酸组成的F-box基序,含有两个高变区(Hva和Hvb)和两个可变区(V1和V2)。生物信息学的方法成功预测了AcSFBd蛋白分子质量为43kD,为亲水性、非分泌型蛋白,在基质内起连接酶(EC6.-.-.-)的作用。成功构建了原核表达载体,并转化大肠杆菌BL21(DE3),SDS-PAGE检测结果显示表达了一个约60kD的蛋白。
A full-length S-hap-lotype-specific F-boxgene (GenBank accession number: FJ362525) was obtained from cultivated almond (Oryza sativa L.) anther in Xinjiang by RT-PCR and RACE techniques and named as AcSFBd. The gene is 1125 bp in length and encodes 374 amino acids. It contains an F-box motif of about 50 amino acids at the N-terminus, containing two hypervariable regions (Hva and Hvb) and two variable regions (V1 and V2) . The bioinformatics method successfully predicts the AcSFBd protein molecular mass of 43kD, is a hydrophilic, non-secreted protein, ligase (EC6.-.-.-) role in the matrix. The prokaryotic expression vector was successfully constructed and transformed into E. coli BL21 (DE3). The result of SDS-PAGE showed that a protein of about 60 kD was expressed.