铁皮石斛DoLEA2基因的克
发布时间：2018-11-02 07:23
【摘要】：晚期胚胎富集蛋白(late embryogenesis abundant proteins,LEA)是一种亲水、热稳定性蛋白,与植物抗逆性密切相关,研究LEA家族基因对铁皮石斛抗逆性研究具有重要意义。本研究以铁皮石斛(Dendrobium officinale)种子为材料,采用RACE技术获得全长cDNA,命名为DoLEA2(Gen Bank登录号:KY626329)。该基因全长cDNA为1 224 bp,编码313个氨基酸,具有晚期胚胎富集蛋白(LEA_2)结构域和水分胁迫高敏感响应结构域,所编码蛋白与其他物种LEA2蛋白具有较高相似性,在进化上与单子叶植物聚为一支,且与蝴蝶兰(Phalaenopsis aphrodite)亲缘关系最近。铁皮石斛DoLEA2在叶的表达量最高,根和茎次之。该基因在Escherichia coli BL21(DE3)中能够高效表达,其优化表达条件为:37℃添加终浓度0.5 mmol·L~(-1)的异丙基-β-D-硫代半乳糖苷(isopropylβ-D-1-thiogalactopyranoside,IPTG)诱导4 h。盐胁迫条件下与对照组相比,重组pET28a-DoLEA2菌株表现出较好的耐盐性。本研究首次获得了铁皮石斛LEA2全长cDNA序列,验证了其耐盐胁迫功能,为进一步研究铁皮石斛抗逆性分子机制提供科学依据。
[Abstract]:Late embryo enriched protein (late embryogenesis abundant proteins,LEA) is a hydrophilic and thermostable protein, which is closely related to plant stress resistance. It is of great significance to study the stress resistance of Dendrobium candidum (Dendrobium candidum) by studying LEA family genes. In this study, (Dendrobium officinale) seeds of Dendrobium candidum were used as materials and RACE technique was used to obtain the full length cDNA, named DoLEA2 (Gen Bank accession number: KY626329). The full-length cDNA of this gene is 1 224 bp, encoding 313 amino acids, with late embryo enriched protein (LEA_2) domain and water stress sensitive domain. The encoded protein is similar to LEA2 protein of other species. The phylogenetic relationship with monocotyledonous plants was similar to that of Phalaenopsis (Phalaenopsis aphrodite). The DoLEA2 expression of Dendrobium candidum was the highest in leaves, followed by roots and stems. The gene was highly expressed in Escherichia coli BL21 (DE3), and the optimal expression conditions were as follows: isopropyl 尾 -D-thiogalactopyranoside (isopropyl 尾 -D-1-thiogalactopyranoside) was added at 37 鈩，
本文编号：2305344
[Abstract]:Late embryo enriched protein (late embryogenesis abundant proteins,LEA) is a hydrophilic and thermostable protein, which is closely related to plant stress resistance. It is of great significance to study the stress resistance of Dendrobium candidum (Dendrobium candidum) by studying LEA family genes. In this study, (Dendrobium officinale) seeds of Dendrobium candidum were used as materials and RACE technique was used to obtain the full length cDNA, named DoLEA2 (Gen Bank accession number: KY626329). The full-length cDNA of this gene is 1 224 bp, encoding 313 amino acids, with late embryo enriched protein (LEA_2) domain and water stress sensitive domain. The encoded protein is similar to LEA2 protein of other species. The phylogenetic relationship with monocotyledonous plants was similar to that of Phalaenopsis (Phalaenopsis aphrodite). The DoLEA2 expression of Dendrobium candidum was the highest in leaves, followed by roots and stems. The gene was highly expressed in Escherichia coli BL21 (DE3), and the optimal expression conditions were as follows: isopropyl 尾 -D-thiogalactopyranoside (isopropyl 尾 -D-1-thiogalactopyranoside) was added at 37 鈩，
本文编号：2305344
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2305344.html
最近更新
教材专著
