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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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A Comparative Study on Mutagenic Effects of Space Flight and Irradiation of γ-rays on Rice 被引量:10
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作者 WEI Li-jun XU Jian-long +5 位作者 WANG Jun-min YANG Qian luo rong-ting ZHANG Ming-xian BAO Gen- liang SUN Ye-qing 《Agricultural Sciences in China》 CAS CSCD 2006年第11期812-819,共8页
The experiment was conducted to study the mutagenic effects of space environment on seedling growth in M1 generation and plant height and heading date in M2 generation in rice. Two types of lines of growth promotion ... The experiment was conducted to study the mutagenic effects of space environment on seedling growth in M1 generation and plant height and heading date in M2 generation in rice. Two types of lines of growth promotion (GP) and growth suppression (GS) were selected from the recombinant inbred lines (RILs) derived from a cross between a sensitive japonica Lemont and a nonsensitive indica Teqing to compare the mutation frequency and mutagenic efficiency for plant height and heading date in the M2 generation. Space environment resulted in 34.9% higher seedling height (SH) in the GP group than in the control, and 39.1% lower in the GS group than in the control, and there was no difference in seed fertility (SF) between the two groups. In M2, mutants of plant height and heading date can be induced by space treatment in both the two groups with lower mutation frequency and mutagenic efficiency in the GP group than in the GS group. There were no significant differences in the physiological damages in M1 between the two groups after γ-rays irradiation treatment. Mutation frequency and mutagenic efficiency of heading date in M2 were higher in the GS group than in the GP group, and the opposite was true for mutagenic efficiency of plant height although the mutation frequency varied between the two groups. The mutation frequencies of plant height and heading date induced by space environment were obviously lower than those by γ-rays irradiation, but the mutagenic efficiency was the opposite for most of the traits. For the GP and GS of seedlings induced by space environment, the GS had higher mutation frequency and mutagenic efficiency for plant height and heading date than the GP in M2. 展开更多
关键词 RICE space environment-induced mutation mutagenic effect physiological damage mutation breeding
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