家蚕丝腺因子SGF-1的基因克隆及序列结构和表达特征与亚细胞定位

发布时间：2018-10-15 08:35

【摘要】：Fox类转录因子在细胞生长、增殖、分化和胚胎发育等过程中发挥重要功能,SGF-1是家蚕丝腺细胞转录因子,属于Fox类转录因子家族。家蚕SGF-1(BGIBMGA005101-TA)基因位于第25号染色体nscaf2823:112743~113792(+strand),含有1个外显子,CDS全长1 050 bp,编码349个氨基酸。家蚕品种大造5龄第3天幼虫的基因芯片表达谱显示,SGF-1基因在大多数组织中有表达,在丝腺中高量表达,其中在前部和中部丝腺的表达量略高于后部丝腺,在雄蚕中的表达量略高于雌蚕。氨基酸组成分析显示,SGF-1蛋白富含脯氨酸、丝氨酸、丙氨酸和亮氨酸,且多数为亲水性氨基酸。序列结构分析表明,SGF-1包含典型的Forkhead结构域,其两端分别为Forkhead_N末端结构域和HNF3_C末端结构域,其中大多数氨基酸残基形成了无规则卷曲结构,α-螺旋与β-折叠结构分别约占18.0%和0.6%。从家蚕后部丝腺克隆了SGF-1基因,并构建麦芽糖结合蛋白标签融合表达载体,表达并纯化了重组SGF-1蛋白。通过RT-PCR与Western blotting检测分别从基因转录水平和蛋白质表达水平上证实,SGF-1在家蚕5龄第3天幼虫丝腺中高量表达。亚细胞定位实验显示SGF-1定位于细胞核中。这些结果为深入研究SGF-1的结构和功能提供了有益的基础信息。
[Abstract]:Fox transcription factors play an important role in cell growth, proliferation, differentiation and embryonic development. SGF-1 is a transcription factor of silkworm gland cells and belongs to the family of Fox transcription factors. The SGF-1 (BGIBMGA005101-TA) gene of Bombyx mori (Bombyx mori) is located on chromosome 25, nscaf2823:112743~113792 (strand), contains one exon, and the CDS length of 1 050 bp, encodes 349 amino acids. The microarray analysis of the 3rd day larva of the 5th instar of silkworm variety showed that the SGF-1 gene was expressed in most tissues and high in the silk gland. The expression of SGF-1 gene in the anterior and middle silk gland was slightly higher than that in the posterior silk gland. The expression level in male silkworm was slightly higher than that in female silkworm. Amino acid composition analysis showed that SGF-1 protein was rich in proline, serine, alanine and leucine, and most of them were hydrophilic amino acids. Sequence analysis showed that SGF-1 contained a typical Forkhead domain, with Forkhead_N terminal domain and HNF3_C terminal domain at both ends. Most of the amino acid residues formed irregularly curled structures, and 伪 -helix and 尾 -fold structures accounted for 18.0% and 0.6%, respectively. The SGF-1 gene was cloned from the posterior silk gland of Bombyx mori and the fusion expression vector of maltose binding protein label was constructed. The recombinant SGF-1 protein was expressed and purified. The results of RT-PCR and Western blotting showed that SGF-1 was highly expressed in the silk gland of silkworm larvae on the 3rd day of 5th instar of silkworm from the level of gene transcription and protein expression. Subcellular localization assay showed that SGF-1 was located in the nucleus. These results provide useful basic information for further study of the structure and function of SGF-1.
【作者单位】：西南大学生物技术学院重庆市蚕丝纤维新材料工程技术研究中心;家蚕基因组生物学国家重点实验室西南大学;
【基金】：国家重点基础研究发展计划“973”项目(No.2012CB1146-02) 国家自然科学基金项目(No.31402139) 中央高校基本科研业务费项目(No.XDJK2013C049;XDJK2013A019) 西南大学博士基金项目(No.SWU112086;SWU112111)
【分类号】：S881.24;Q78

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