Hubei University

Wuhan Research Institute of Materials Protection

Hubel University

Lixin Ma

Fei Wang

Yang Liu

Wanping Chen

Haitao Duan

Dan Jia

Shengpeng Zhan

Tian Yang

Jiesong Tu

Yinhua Li

Chao Zhai

Xiaochen Xie

Longyu Wang

Yu Wang

Jian Li

Jun Yang

Shihui Yang

Rey-Ting Guo

Chun-Chi Chen

Jian-Wen Huang

Longhai Dai

Lilan Zhang

Xuan Yang

Wulin Zhang

Wei Shen

Yuan Wang

Qi Liu

Xiaoman Jiang

Yijie Jin

Fuyong Huang

Yanming Dong

Junbo Cao

Runxia Li

Xuejing Yu

Jiakai Cui

Miaomiao Chen

Yaping Wang

Chin-Yu Chen

Zhibin Fan

Yibin Cheng

Ju Huang

Shan Wu

Xingeng Li

Zhichun Zhan

Yunhu Ding

Yongfu Yang

Li Yi

Wenqiang Li

Ruyi He

Provided are a recombinase polymerase amplification (RPA)-PAND based enterovirus virus typing detection kit and a rapid sensitive detection method, where the detection kit includes a PfAgo protein three gDNAs(g1/g2/g3), MB and its corresponding gDNA(gMB) combination for specifically detection of enteroviruses EV71, CVA6, CVA10, CVA16 and the universal enterovirus (EVU). RPA-HFMD-PAND rapid nucleic acid detection methods for typing detection of common enterovirus type A pathogens EV71, CVA6, CVA10, CVA16 and enterovirus universal EVU are successfully established based on PfAgo protein mediated target detection in combination with an RPA technology. The method has the advantages of high sensitivity, high specificity, low cost, relatively low position targeting restriction, simple reaction system and no aerosol pollution, and can conduct multi-channel nucleic acid detection. The present disclosure provides a new reagent and approach for clinic diagnosis and epidemiology monitoring of a hand-foot-mouth disease (HFMD).

RPA-PAND based enterovirus typing detection kit and detection method

Argonaute proteins from prokaryotes (pAgos) and applications thereof are provided, the pAgo is from a mesophilic prokaryote ofand named as KmAgo, the pAgos and applications thereof are capable of site-specific modification of intracellular and extracellular genetic material, and thus can be effectively applied to many fields of biotechnology, such as nucleic acid detection, gene editing and gene modification, and provide a new tool for gene editing, modification and molecular detection of the pAgo polypeptides.

Argonaute proteins from prokaryotes and applications thereof

An efficient synthesis and assembly method for the large fragment DNA based on the programmable nuclease Argonaute, specifically includes: constructing and treating antibiotic resistance gene reconstructed vectors with linearization, dividing a target DNA into multiple small DNA fragments and then synthesizing the small DNA fragments, followed by loading the synthesized small DNA fragments to the antibiotic resistance gene reconstructed vectors; and the SLIC and resistance gene reconstruction are used to achieve assembly of the target DNA. The method combines the SLIC with a resistance gene reconstruction strategy, allowing for the assembly of 5-6 small fragments in a single resistance gene reconstruction, which is more efficient and time-saving. Moreover, the number of the resistance gene reconstructions can be flexibly chosen according to the length of the DNA fragments. Mutations are not introduced caused by PCR, and the reconstructed large fragment do not need a second sequencing, saving time and costs.

Efficient synthesis and assembly method for large fragment DNA based on programmable nuclease Argonaute

AArgonaute (PfAgo) mutant protein with mesophilic target nucleic acid cleavage activity and an application thereof are provided. The PfAgo mutant protein has an amino acid mutation on at least one of a 617position and a 618position relative to the sequence shown in SEQ ID NO: 1, specifically, K617E/G and/or L618Y/F/W/G. Compared with a wild-type PfAgo protein, the provided PfAgo mutant protein has activity in the temperature range of 30-95 Celsius degree (° C.), effectively expanding application range of prokaryotic Argonaute (pAgo) protein.

PfAgo mutant protein mesophilic target nucleic acid cleavage activity and application thereof

An application of a prokaryotic Argonaute protein with only a cleavage activity of a target ribonucleic acid (RNA) in RNA editing is provided. The Argonaute protein is derived from a mesophilic prokaryotes, and its amino acid sequence is shown in SEQ ID NO: 1 or a protein with high similarity to SEQ ID NO: 1 and the same function. This protein has binding activity to a single-stranded guide DNA and nuclease activity only to the target RNA complementarily paired with the single-stranded guide DNA. Therefore, the protein can be utilized for in vitro and in vivo targeted RNA editing, providing a new powerful tool for RNA editing.

Application of prokaryotic Argonaute protein with only RNA target cleavage activity in RNA editing

A titanium dioxide micro-nanocontainers, corrosion-resistant waterborne epoxy coatings and preparation method thereof, including preparation steps as follows: TiOmicro-nano spheres are synthesized by applying hydrothermal method; a polyaniline layer doped with molybdate ions is deposited on the surface of TiOmicro-nano spheres by adopting the method of in-situ chemical polymerization, TiO/PANI-MoOmicro-nano-spheres are obtained, then, polydopamine is encapsulated on the surface of TiO/PANI-MoOmicro-nano spheres to obtain titanium dioxide micro-nanocontainers; next, antirust filler, defoamer, dispersant and thickener are added into waterborne epoxy emulsion, then titanium dioxide micro-nanocontainers are added in the waterborne epoxy emulsion for dispersing and grinding, filtering and encapsulating to obtain component A; the waterborne epoxy curing agent and deionized water are mixed in proportion to obtain component B; component A is stirred, then it is mixed with the component B in proportion, corrosion-resistant waterborne epoxy coatings is obtained. According to the invention, the titanium dioxide micro-nanocontainers is synthesized and added into the coating as an additive, which can not only improve the compatibility between the filler and the emulsion, but greatly improves the long-term corrosion resistance of the coating by prolonging the release time of the corrosion inhibitors.

Titanium dioxide micro-nanocontainers, corrosion-resistant waterborne epoxy coating and preparation method thereof

A protein complex based on DNA enzymes of an E family ofand an application thereof in artificial protein scaffolds are provided. The protein complex includes one or more of interaction pairs formed by a CL2 protein and an Im2 protein, a CL7 protein and an Im7 protein, a CL8 protein and an Im8 protein, or a CL9 protein and an Im9 protein. By protein engineering of a carboxyl terminus DNase domain of the DNA enzymes CE2, CE7, CE8 and CE9, mutants that lose DNA enzyme activity but still retain the ultra-high affinity with the corresponding Im protein are obtained, and protein interaction pairs CL2/Im2, CL7/Im7, CL8/Im8 and CL9/Im9 are constructed. These protein interaction pairs have properties of heat resistance, high affinity, high specificity, small molecular weight, fast assembly speed, etc. Based on this, an artificial protein scaffold is constructed for the construction of artificial multienzyme complexes.

Protein complex based on DNA enzymes of E family ofand application thereof in artificial protein scaffolds

Mbp_Argonaute proteins from prokaryotes and application thereof are provided. The Mbp_Argonaute protein consists of an amino acid sequence as shown in SEQ ID NO: 1 or a sequence with at least 50% or at least 80% of homology with the amino acid sequence as shown in SEQ ID NO: 1. An Argonaute protein gene derived from a cold-resistant prokaryoteis synthesized and named as MbpAgo, which has binding activity to single-stranded guide DNA and has nuclease activity to target RNA and/or target DNA complementarily paired with the single-stranded guide DNA, the MbpAgo can be used for the target RNA editing in vivo and in vitro to achieve site-specific modification of genetic material. The MbpAgo can modify highly-structured RNAs and not affect an endogenous RNAi pathway of animal and plant cells, provides a new and powerful tool for RNA editing with high cleavage activity and good specificity.

Mbp_Argonaute proteins from prokaryotes and applications thereof

A graphene reinforced aluminum matrix composite with high electrical conductivity and a preparation method thereof. The method includes: obtaining aluminum coated graphene powder by plating aluminum on a graphene surface, melting aluminum block into aluminum liquid, heating a mold to be lower than an aluminum melting point, alternately pouring the aluminum liquid and the aluminum coated graphene powder into the mold for layered casting to obtain a sandwich structure; extruding the sandwich structure into a rectangular test block and then heating to 500&#x2dc;600&#xb0; C., performing heat preservation for a preset time and performing forging treatment, and performing longitudinal cold deformation under inert gas to obtain the graphene reinforced aluminum matrix composite. The method can solve a problem that poor wettability of graphene and aluminum matrix, the graphene is evenly dispersed in the aluminum matrix, which can improve strength of the aluminum matrix and keep its high electrical conductivity.

Graphene reinforced aluminum matrix composite with high electrical conductivity and preparation method thereof

A method for identifying and characterizing biological parts based on omics datasets and a dual-fluorescent reporter gene system, and a biological part library constructed thereon are provided, relating to a technical filed of biology. The method includes steps of: identifying the biological parts using the omics datasets; constructing a single-fluorescent reporter gene system using a shuttle vector pEZ15Asp as a skeleton for screening and determining fluorescent reporter genes; obtaining a dual-fluorescent reporter gene system skeleton; constructing recombinant plasmids, and finally transforming into competent cells for quantitative analysis of fluorescence intensities. The present invention is convenient and quick, and can screen and identify different biological parts such as RBS, UTRs, promoters, and terminators of different intensities in batch quantitatively in a relatively short time. Moreover, the present invention can quickly expand the biological part library of, so as to be applied in metabolic engineering of different demands.

Method for characterizing biological part based on dual-fluorescent reporter gene system and biological part library constructed thereon

A cellulase having improved enzymatic activity is disclosed. The cellulase has a modified amino acid sequence of SEQ ID NO: 2 or a modified amino acid sequence with at least 80% sequence identity of SEQ ID NO: 2, wherein the modification is a substitution of methionine at position 120 or a corresponding position with asparagine.

Cellulase having improved enzymatic activity

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Wuhan, China

Lixin Ma