期刊论文

中文

湖南农业大学学报(自然科学版)

1007-1032

2022

48

3

294-297

10.13331/j.cnki.jhau.2022.03.007

科学技术部国家重点研发计划（2021YFD1400800）； 湖南省科学技术厅重点研发计划项目（2022NK2052）； 湖南农业大学双一流学科建设项目（SYL2019027）；

本校为第一机构

为进一步了解柑橘黄龙病菌亚洲种Clas的致病机理，从Prasad预测的166种分泌蛋白中选取1个在柑橘植株中表达量较高的CLIBASIA–04580基因进行研究。提取感染柑橘黄龙病的柑橘叶片总DNA，经 PCR扩增得到 CLIBASIA–04580基因，切胶回收PCR产物双酶切连接到pET–28a载体上，使用菌落PCR和限制性内切酶双酶切鉴定筛选阳性克隆；转化大肠杆菌 BL21（DE3），使用0.5、1.0、1.5、2.0 mmol/L的异丙基硫代半乳糖苷（IPTG） 诱导重组蛋白表达融合组氨酸标签的CLIBASIA–04580重组蛋白，聚丙烯酰氨凝胶电泳（SDS–PAGE）检测重组蛋白的表达水平，然后使用蛋白纯化镍柱进行纯化。结果表明，携带组氨酸标签的...

CLIBASIA-04580,which highly expressed in citrus plants,was selected from 166 secreted proteins predicted by Prasad and was investigated to further understand the pathogenesis of Candidatus Liberibacter asiaticus(CLas).The total DNA of citrus leaves identified as infected with citrus Huanglongbing(HLB)was extracted,and the CLIBASIA-04580 gene was obtained by PCR.The PCR product was extracted and connected to the pET-28αvector by double enzyme digestion,and the positive clones were screened by colony PCR and restriction endonuclease double diges...