特早熟春性甘蓝型油菜sBnFLD基因的克隆及表达
发布时间：2018-09-05 11:17
【摘要】：【目的】克隆特早熟春性甘蓝型油菜A基因组上的sBnFLD基因,并对其进行表达研究,为该基因的功能及其在成花转变中的作用研究奠定基础。【方法】根据GenBank中已报道的拟南芥和白菜型油菜FLD同源基因的保守序列设计引物,采用PCR和RT-PCR扩增特早熟春性甘蓝型油菜86号品系(光周期不敏感)的FLD同源基因,用qRT-PCR技术检测sBnFLD基因在86号品系不同发育时期茎、叶和茎尖中的表达情况。【结果】克隆出了sBnFLD基因,命名为sBnFLD,在GenBank中的登录号为KR003079.1。sBnFLD基因cDNA全长2 376bp,有3个内含子,4个外显子,编码791个氨基酸残基,分子量86.5ku,等电点8.5;sBnFLD为非分泌蛋白和非膜蛋白;sBnFLD蛋白N端有2个保守的结构域α螺旋结构域(SWIRM)和NAD(P)-binding-8结构域,该蛋白有多个α螺旋和β折叠。生物信息学分析显示,sBnFLD蛋白与已报道的甘蓝型油菜未知蛋白(CDX73929.1)和电子克隆的白菜型油菜FLD(XP_009135110.1)氨基酸序列相似性达99%,与拟南芥FLD氨基酸序列相似性达87%。qRT-PCR分析结果显示,sBnFLD基因在油菜苗期和现蕾初期茎、叶、茎尖中均有表达,但在蕾期茎尖中表达量最高。【结论】克隆出的sBnFLD基因为甘蓝型油菜的FLD同源基因,该基因在春性特早熟甘蓝型油菜开花调控中可能起着重要的调节作用。
[Abstract]:[objective] to clone and express sBnFLD gene from A genome of especially precocious vernal Brassica napus L. [methods] Primer was designed according to the conserved sequence of FLD homologous gene of Arabidopsis thaliana and Brassica campestris reported in GenBank. PCR and RT-PCR were used to amplify the FLD homologous gene of rape 86 (photoperiod insensitive), and the sBnFLD gene was detected by qRT-PCR technique in stem of 86 strain at different developmental stages. The expression of sBnFLD gene in leaf and stem tip was cloned. The accession number of sBnFLD, in GenBank was 2376 BP of KR003079.1.sBnFLD gene, with 3 introns and 4 exons, encoding 791 amino acid residues. The molecular weight is 86.5ku.The isoelectric point 8.5 sBnFLD is a non-secretory protein and a non-membrane protein. There are two conserved domains, 伪 helix (SWIRM) and NAD (P) binding-8, which have multiple 伪 helix and 尾 folding. Bioinformatics analysis showed that the amino acid sequence similarity of BnFLD protein with unknown protein (CDX73929.1) and FLD (XP_009135110.1) sequence of Brassica napus and Arabidopsis thaliana FLD was 990.The result of 87%.qRT-PCR analysis showed that the amino acid sequence was similar to that of Arabidopsis thaliana FLD. SBnFLD gene was used in the stem of rapeseed at seedling stage and early budding stage. Both leaf and stem tip were expressed, but they were the highest in bud stage. [conclusion] the cloned sBnFLD gene was the FLD homologous gene of Brassica napus. This gene may play an important role in the regulation of flowering in spring especially early maturing Brassica napus.
【作者单位】： 青海大学生态环境工程学院;青海大学农林科学院;
【基金】：国家自然科学基金项目(31360345) 青海省科技厅项目(2016-ZJ-787) 国家“863计划”项目(2011AA10A104) 科技部科技支撑计划项目(2013BAD01B05-3)
【分类号】：S565.4
，
本文编号：2224110
[Abstract]:[objective] to clone and express sBnFLD gene from A genome of especially precocious vernal Brassica napus L. [methods] Primer was designed according to the conserved sequence of FLD homologous gene of Arabidopsis thaliana and Brassica campestris reported in GenBank. PCR and RT-PCR were used to amplify the FLD homologous gene of rape 86 (photoperiod insensitive), and the sBnFLD gene was detected by qRT-PCR technique in stem of 86 strain at different developmental stages. The expression of sBnFLD gene in leaf and stem tip was cloned. The accession number of sBnFLD, in GenBank was 2376 BP of KR003079.1.sBnFLD gene, with 3 introns and 4 exons, encoding 791 amino acid residues. The molecular weight is 86.5ku.The isoelectric point 8.5 sBnFLD is a non-secretory protein and a non-membrane protein. There are two conserved domains, 伪 helix (SWIRM) and NAD (P) binding-8, which have multiple 伪 helix and 尾 folding. Bioinformatics analysis showed that the amino acid sequence similarity of BnFLD protein with unknown protein (CDX73929.1) and FLD (XP_009135110.1) sequence of Brassica napus and Arabidopsis thaliana FLD was 990.The result of 87%.qRT-PCR analysis showed that the amino acid sequence was similar to that of Arabidopsis thaliana FLD. SBnFLD gene was used in the stem of rapeseed at seedling stage and early budding stage. Both leaf and stem tip were expressed, but they were the highest in bud stage. [conclusion] the cloned sBnFLD gene was the FLD homologous gene of Brassica napus. This gene may play an important role in the regulation of flowering in spring especially early maturing Brassica napus.
【作者单位】： 青海大学生态环境工程学院;青海大学农林科学院;
【基金】：国家自然科学基金项目(31360345) 青海省科技厅项目(2016-ZJ-787) 国家“863计划”项目(2011AA10A104) 科技部科技支撑计划项目(2013BAD01B05-3)
【分类号】：S565.4
，
本文编号：2224110
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2224110.html
最近更新
教材专著
