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抗菌肽CM4基因克隆及其在毕赤酵母中的表达鉴定 总被引:7,自引:0,他引:7
为了在毕赤酵母中获得抗菌肽CM4表达,将抗菌肽CM4基因克隆入表达质粒pPIC9,SacⅠ线性化的重组表达质粒pPIC9-CM4电击转化P.pastorisGS115(his-),采用MM、MD板和PCR法筛选Mut 表型,甲醇诱导表达;抗菌活性检测、Tricine-SDS-PAGE及酸性活性电泳均表明抗菌肽CM4在毕赤酵母中成功地分泌表达;重组抗菌肽CM4对黑曲霉(Aspergillus niger)、绿色木霉(Trichoderma viride)都有很强的抑菌活性。 相似文献
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参照天然抗菌肽CM4(ABP-CM4)氨基酸序列和大肠杆菌偏爱密码子,采用rPCR法获得CM4基因后重组到表达载体pET32a上,在E.coli中融合表达。表达产物以可溶性存在,经Ni2 -NTA琼脂糖亲和层析获得融合蛋白,再经甲酸切割、亲和层析和阳离子交换层析,得到纯化的重组抗菌肽。琼脂糖扩散法和液相测定法证明了纯化的抗菌肽具有抗菌活性。 相似文献
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From 2009 to 2020, we systematically investigated the population, mortality, and death factors of Père David’s deer (Elaphurus davidianus) in the Dafeng Milu National Nature Reserve, Jiangsu, China. The result showed the annual growth rate of the population was 13.09% ± 5.96%, which continuously increased in the 12 years. The annual mortality rate was 3.53% ± 2.27%, with an obvious fluctuation every 2 years. There was no significant difference in mortality of adults and sub-adults in different seasons, but significant in juveniles.Significant differences in mortality among different age groups were observed. The adults died most in summer and autumn, while the juveniles died most in winter and spring. 48.77% of subjects tend to choose the underbrush for dying. The death factors among different age groups were significantly different. The elders and the juveniles mainly died of epidemic diseases, including viral, bacterial, and parasitic. While the sub-adults died in accidents. We suggested that it is necessary to strengthen the daily monitoring and epidemic disease prevention in wildlife management. Our data provided important information for the protection of Père David’s deer in Dafeng Reserve. 相似文献
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