狗头枣GSTU基因的克隆及其序列分析

Cloning and Sequence Analysis of GSTU Gene from Goutou Jujube

为探索红枣GSTU类基因在红枣抗逆中的分子作用机制,本研究以狗头枣枣树叶片的DNA为模板,根据枣GSTU基因的序列(HM345954.1)设计引物,采用PCR方法获得GSTU基因序列,并利用生物信息学手段对其所对应蛋白的结构域、功能域、理化特性、跨膜区、二级结构及亚细胞定位等进行分析。结果表明:克隆到的DNA序列全长838 bp,具有2个外显子和1个内含子,内含子在292~460 bp碱基处,该序列与GenBank中枣的GSTU核苷酸序列和氨基酸序列的同源性分别为99.85%和100%,表明成功的克隆了狗头枣的GSTU基因。该序列编码222个氨基酸,分子量为25.268 kD,理论等电点(pI)为6.10;亲水性的平均数为-0.242,表明该蛋白是亲水性蛋白。二级结构主要以α-螺旋为主,其次是无规则卷曲,不存在信号肽,表明该蛋白可能为非分泌性蛋白,该蛋白可能位于内质网(膜)和质膜上。本研究获得的狗头枣GSTU基因在基因结构上含有1个内含子,在核酸序列及氨基酸序列上与GenBank中枣的GSTU基因同源性高,并对其理化性质、结构等进行了预测与分析,为进一步研究红枣抗逆分子机制提供理论基础,同时为红枣品种的选育提供理论支持。

In order to explore the molecular mechanism of GSTU genes in the stress resistance of data, this study selects DNA in the leaves of a kind of Chinese data called Goutouzao as sample, and primers were designed according to the sequence of GSTU gene(HM345954.1), which was obtained by PCR. The structural domain, functional domain, physicochemical properties, transmembrane region, secondary structure and subcellular localization of the corresponding protein were analyzed by bioinformatics. The results showed that the cloned DNA sequence was 838 bp in length, with 2 exons and 1 intron. The intron was located at 292~460 bp bases, and the homology of the sequence with the GSTU nucleotide sequence and amino acid sequence in GenBank was 99.85% and 100%, respectively, indicating that the GSTU gene of jujube was successfully cloned. The sequence encoded 222 amino acids with a molecular weight of 25.268 kD and a theoretical isoelectric point(pI) of 6.10;The average hydrophilicity was-0.242, indicating that the protein was hydrophilic. The secondary structure is dominated by alpha-helix, followed by random coil, and there is no signal peptide, indicating that the protein may be non-secretory, and that the protein may be located in the endoplasmic reticulum(membrane) and plasma membrane. The GSTU gene of Goutouzao obtained in this study contains a introns genetic structure, on nucleic acid and amino acid sequences in GenBank data GSTU gene homology is high, and the author also predicted and analyzed its physicochemical properties, structure and so on to provide the theoretical basis for further studies in this field and provide theoretical support for the breeding of red jujube varieties at the same time.