期刊论文

Preparation and application of rabbit-derived polyclonal antibody against fatty acid desaturase 2 in Brassica napus

中文

江苏农业学报

1000-4440

2020

36

04

851-857

10.3969/j.issn.1000-4440.2020.04.007

国家转基因生物新品种培育科技重大专项（2018ZX08011-01B）； 湖南省重点领域研发计划项目（2019DK2011）；

本校为第一机构

脂肪酸脱氢酶2(FAD2)是调控油菜种子油酸含量的关键酶。为建立其灵敏、便捷的免疫学检测方法,本研究合成了BnFAD2(UniProtKB:C3W518)中2个多肽片段,与KLH载体蛋白偶联,混合免疫新西兰大白兔,分离纯化得到了油菜BnFAD2兔源多克隆抗体。应用于FAD2-RNAi高油酸转基因油菜品系W-4不同世代果荚及成熟叶片总蛋白质的免疫印迹检测,结果表明,该兔源多克隆抗体,可在油菜内质网成分中识别FAD2,特征性条带大小为95 000。在果荚总蛋白质样本中,WT的FAD2含量高于转基因材料W-4,而在叶片总蛋白质样本中WT和W-4的FAD2含量相同。本研究成功制备出BnFAD2兔源多克隆抗体,并首次验证出油菜内源性FAD2以多聚体形式行...

Fatty acid desaturase 2(FAD2)is the key enzyme regulating oleic acid content.The immunological methods for its rapid detection are limited by the lack of antibodies.In this study,the Brassica napus FAD2 polyclonal antibodies were successfully prepared with the rabbit immunized by two synthesized polypeptides from BnFAD2(UniProtKB: C3W518).The pods or mature leaves extracts of different generations of W-4 which was the RNAi transgenic material were detected by western blot.The results showed that the rabbit-derived polyclonal antibody could recognize the FAD2 in the endoplasmic reticulum,and th...