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Protein purification is the vital basis to study the function, structure and interaction of proteins. Widely used methods are affinity chromatography-based purifications, which require different chromatography columns and harsh conditions, such as acidic pH and/or adding imidazole or high salt concentration, to elute and collect the purified proteins. Here we established an easy and fast purification method for soluble proteins under mild conditions, based on the light-induced protein dimerization system improved light-induced dimer (iLID), which regulates protein binding and release with light. We utilize the biological membrane, which can be easily separated by centrifugation, as the port to anchor the target proteins. In Xenopus laevis oocyte and Escherichia coli, the blue light-sensitive part of iLID, AsLOV2-SsrA, was targeted to the plasma membrane by different membrane anchors. The other part of iLID, SspB, was fused with the protein of interest (POI) and expressed in the cytosol. The SspB-POI can be captured to the membrane fraction through light-induced binding to AsLOV2-SsrA and then released purely to fresh buffer in the dark after simple centrifugation and washing. This method, named mem-iLID, is very flexible in scale and economic. We demonstrate the quickly obtained yield of two pure and fully functional enzymes: a DNA polymerase and a light-activated adenylyl cyclase. Furthermore, we also designed a new SspB mutant for better dissociation and less interference with the POI, which could potentially facilitate other optogenetic manipulations of protein–protein interaction. 相似文献
53.
Ruiqi Liu Tingting Chen Xiao Yin Gaoqing Xiang Jing Peng Qingqing Fu Mengyuan Li Boxing Shang Hui Ma Guotian Liu Yuejin Wang Yan Xu 《The Plant journal : for cell and molecular biology》2021,106(6):1557-1570
Pathogens secrete a large number of effectors that manipulate host processes to create an environment conducive to pathogen colonization. However, the underlying mechanisms by which Plasmopara viticola effectors manipulate host plant cells remain largely unclear. In this study, we reported that RXLR31154, a P. viticola RXLR effector, was highly expressed during the early stages of P. viticola infection. In our study, stable expression of RXLR31154 in grapevine (Vitis vinifera) and Nicotiana benthamiana promoted leaf colonization by P. viticola and Phytophthora capsici, respectively. By yeast two-hybrid screening, the 23-kDa oxygen-evolving enhancer 2 (VpOEE2 or VpPsbP), encoded by the PsbP gene, in Vitis piasezkii accession Liuba-8 was identified as a host target of RXLR31154. Overexpression of VpPsbP enhanced susceptibility to P. viticola in grapevine and P. capsici in N. benthamiana, and silencing of NbPsbPs, the homologs of PsbP in N. benthamiana, reduced P. capcisi colonization, indicating that PsbP is a susceptibility factor. RXLR31154 and VpPsbP protein were co-localized in the chloroplast. Moreover, VpPsbP reduced H2O2 accumulation and activated the 1O2 signaling pathway in grapevine. RXLR31154 could stabilize PsbP. Together, our data revealed that RXLR31154 reduces H2O2 accumulation and activates the 1O2 signaling pathway through stabilizing PsbP, thereby promoting disease. 相似文献
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平须舞虻属Platypalpus Macquart是舞虻科中最大的属之一,已知550余种分布世界各地.四川以前已知8种.本文记述四川平须舞虻属3新种:短芒平须舞虻Platypalpus breviarista sp.nov.,微距平须舞虻Platypalpus minor sp.nov.和亚丁平须舞虻Platypalpus yadinganus sp.nov,均属于长角平须舞虻种团Platypalpus longicornis species group,并编制了四川平须舞虻属分种检索表. 相似文献
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Zhu Yun Xu Baoping Li Changchong Chen Zhimin Cao Ling Fu Zhou Shang Yunxiao Chen Aihuan Deng Li Bao Yixiao Sun Yun Ning Limin Yu Shuilian Gu Fang Liu Chunyan Yin Ju Shen Adong Xie Zhengde Shen Kunling 《中国病毒学》2021,36(6):1543-1553
Virologica Sinica - Community-acquired pneumonia (CAP) is one of the leading causes of morbidity and mortality in children worldwide. In this study, we aimed to describe the aetiology of viral... 相似文献
57.
正In 2013, tumor immunotherapy topped the list of the top ten scientific breakthroughs(Couzin-Frankel, 2013), and it was widely used in the treatment of lung cancer, kidney cancer,and melanoma(Routy et al., 2018). However, the response rate of patients to tumor immunotherapy varies, and usually,only a small percentage of patients respond well to treatment(Sambi et al., 2019). 相似文献
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Xingdong Zhou Hui Wang Qun Ji Mingjuan Du Yuexia Liang Huanhuan Li Fan Li Hang Shang Xiujuan Zhu Wei Wang Lichun Jiang Alexey V.Stepanov Tianyu Ma Nanxin Gong Xiaodong Jia Alexander G.Gabibov Zhiyong Lou Yinying Lu Yu Guo Hongkai Zhang Xiaoming Yang 《蛋白质与细胞》2021,12(10):818-823
Dear Editor,
The rapid emergence and persistence of the pandemic caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) has had enormous impacts on global health and the economy.Effective vaccines against SARS-CoV-2 are urgently needed to control the coronavirus disease 2019(COVID-19) pandemic,and multiple vaccines have been found to be efficacious in preventing symptomatic COVID-19(Polack et al.,2020;Wu et al.,2020;Jones and Roy,2021).We have developed a traditional beta-propiolactone-inacti-vated aluminum hydroxide-adjuvanted whole-virion SARS-CoV-2 vaccine (BBIBP-CorV),which elicited protective immune responses in clinical trials (Wang et al.,2020;Xia et al.,2021).The vaccine has been granted conditional approvals or emergency use authorizations (EUAs) in China and other countries. 相似文献
59.
Liao Jingqiu Cai Yan Wang Xinrui Shang Chenxu Zhang Qian Shi Huizhong Wang Shifeng Zhang Dongdong Zhou Yongcan 《Probiotics and antimicrobial proteins》2021,13(4):1119-1137
Probiotics and Antimicrobial Proteins - A potential host-derived probiotic, Bacillus subtilis 6-3-1, was successfully screened from 768 isolates from the intestines of healthy hybrid grouper... 相似文献
60.
Wang Che Huang Lili Li Ruojin Wang Ying Wu Xiaoxue Shang Dejing 《International journal of peptide research and therapeutics》2021,27(4):2291-2301
International Journal of Peptide Research and Therapeutics - Multidrug resistance (MDR) is one of the major obstacles to efficient chemotherapy against cancers, resulting from the overexpression of... 相似文献