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鸭疫里默氏杆菌AS87-01740基因生物信息学分析及其原核表达
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作者 单敏 李涛 +4 位作者 何晶 李文海 王小兰 罗廷荣 于圣青 《南方农业学报》 CAS CSCD 北大核心 2017年第11期2058-2063,共6页
【目的】了解鸭疫里默氏杆菌AS87-01740基因分子生物学特性,确定其编码蛋白抗原性,为鸭疫里默氏杆菌亚单位疫苗研发提供新的靶标抗原选择。【方法】采用PCR克隆鸭疫里默氏杆菌AS87-01740基因,以DNASTAR和NCBI数据库分别进行基因生物信... 【目的】了解鸭疫里默氏杆菌AS87-01740基因分子生物学特性,确定其编码蛋白抗原性,为鸭疫里默氏杆菌亚单位疫苗研发提供新的靶标抗原选择。【方法】采用PCR克隆鸭疫里默氏杆菌AS87-01740基因,以DNASTAR和NCBI数据库分别进行基因生物信息学分析;利用p Cold-TF原核表达载体在大肠杆菌BL21感受态细胞中进行诱导表达,并分别以SDS-PAGE和Western blotting检测融合蛋白的表达形式及其抗原特异性。【结果】从鸭疫里默氏杆菌基因组扩增获得的AS87-01740基因编码框全长618 bp,编码205个氨基酸,理论分子量约21.87 kD,理论等电点(p I)8.2,正电荷氨基酸总数17个,负电荷氨基酸总数16个。AS87-01740蛋白序列在同种属分离株中,与2型血清型分离株(11845、YM、GD和CH-2)有很高的相似性(>99%),与1型血清型分离株(CH-1和CH-3)的相似性为92%;在不同种属中,与金黄杆菌(Chryseobacterium sp.)、动物溃疡伯格菌(Bergeyella zoohelcum)和脑膜炎败血伊丽莎白菌(Elizabethkingia meningosepticum)的相似性分别为61%、60%和57%。经异丙基硫代-β-D半乳糖苷(IPTG)诱导表达获得的融合蛋白分子量约77 kD,以可溶性表达形式为主。Western blotting检测结果表明,融合蛋白能与鸭疫里默氏杆菌阳性血清发生特异性免疫反应。【结论】AS87-01740基因编码蛋白在不同鸭疫里默氏杆菌分离株中具有很高的保守性,且能与其阳性血清发生特异性免疫反应,是研发鸭疫里默氏杆菌疫苗的潜在抗原。 展开更多
关键词 鸭疫里默氏杆菌 as87-01740基因 生物信息学 原核表达 免疫原性
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Differences of aroma development and metabolic pathway gene expression between Kyoho and 87-1 grapes 被引量:6
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作者 JI Xiao-hao W ANG Bao-liang +3 位作者 WANG Xiao-di W ANG Xiao-long LIU Feng-zhi WANG Hai-bo 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2021年第6期1525-1539,共15页
Aroma is an important quality trait of grapes and often the focus of consumers,viticulturists and grapevine breeders.Kyoho is a hybrid between Vitis vinifera and Vitis labrusca with a strawberry-like scent,while 87-1 ... Aroma is an important quality trait of grapes and often the focus of consumers,viticulturists and grapevine breeders.Kyoho is a hybrid between Vitis vinifera and Vitis labrusca with a strawberry-like scent,while 87-1 is an early-ripening mutant of Muscat hamburg,belonging to Vitis vinifera,with a rose scent.In this study,we compared their aroma compositions and concentrations during berry development by headspace-SPME combined with gas chromatography-mass spectrometry(GC-MS),and analyzed the expression differences of enzyme-encoding genes in the LOX-HPL,MEP and MVA metabolic pathways by qRT-PCR.Twelve esters were detected in Kyoho during the whole berry development and they were abundant after veraison,but no esters were detected in 87-1 berries.Linalool was the dominant terpene among the 14 terpenes detected in 87-1 berries,while limited amounts of terpenes were detected in Kyoho berries.qRT-PCR analysis indicated that the low expression of VvAAT might explain the low content of ester volatiles in 87-1 berries,and the low expression of coding genes in the MEP pathway,especially VvPNLin Ner1,might be the reason for the low content of volatile terpenes in Kyoho berries.The results from this work will promote our understanding of aroma metabolic mechanisms of grapes,and offer some suggestions for grape aromatic quality improvement. 展开更多
关键词 GRAPE Kyoho 87-1 AROMA LOX-HPL MEP gene expression
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