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Yeast surface display (YSD) is a technology that fuses the exogenous target protein gene sequence with a specific vector gene sequence, followed by introduction into yeast cells. Subsequently, the target protein is expressed and localized on the yeast cell surface by using the intracellular protein transport mechanism of yeast cells, whereas the most widely used YSD system is the α-agglutinin expression system. Yeast cells possess the eukaryotic post-translational modification mechanism, which helps the target protein fold correctly. This mechanism could be used to display various eukaryotic proteins, including antibodies, receptors, enzymes, and antigenic peptides. YSD has become a powerful protein engineering tool in biotechnology and biomedicine, and has been used to improve a broad range of protein properties including affinity, specificity, enzymatic function, and stability. This review summarized recent advances in the application of YSD technology from the aspects of library construction and screening, antibody engineering, protein engineering, enzyme engineering and vaccine development.
Subject(s)
Saccharomyces cerevisiae/metabolism , Protein Engineering , Biotechnology , Antibodies/metabolism , Amino Acid SequenceABSTRACT
As central players in cellular structure and function, proteins have long been central themes in life science research. Analyzing the impact of protein sequence variation on its structure and function is one of the important means to study proteins. In recent years, a technology called deep mutational scanning (DMS) has been widely used in the field of protein research. It introduces thousands of mutations in parallel in specific regions of proteins through high-abundance DNA libraries. After screening, high-throughput sequencing is employed to score each mutation, revealing sequence-function correlations. Due to its high-throughput, fast and easy, and labor-saving features, DMS has become an important method for protein function research and protein engineering. This review briefly summarizes the principle of DMS technology, highlighting its applications in mammalian cells. Moreover, this review analyzes the current technical bottlenecks, aiming to facilitate relevant research.
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Animals , Mutation , Proteins/chemistry , Protein Engineering , High-Throughput Nucleotide Sequencing/methods , Mammals/geneticsABSTRACT
Objective@#The objective of this study was to establish a next generation sequencing (NGS) method for severe fever with thrombocytopenia syndrome virus(SFTS).@*Methods@#SFTS virus RNA was extracted from the patient serum inoculated and isolated by Vero cells. Two methods of random primer sequencing and oligo(dT) beads selection sequencing were used for library construction. The libraries were built based on the best amplification and purification conditions. Whole genome sequencing was performed on NGS platform.@*Results@#There were significant differences in data of 3 virus between the two methods.The sample was sequenced by random primer sequencing showed low coverage and depth. However, three samples were sequenced by oligo(dT) beads selection showed coverage was over 99% and depth was over 900.The alignment rate of database from NCBI was more than 90%. The initial detection quality of this method was 300ng RNA.@*Conclusions@#In this study, we used the method of oligo(dT) beads selection to build libraries, and established a SFTS virus genome detection based on next generation sequencing.
ABSTRACT
OBJECTIVE@#To optimize DNA library construction in non-crosslinked chromatin immunoprecipitation coupled with next-generation sequencing (Native ChIP-seq) to obtain high-quality Native ChIP-seq data.@*METHODS@#Human nasopharyngeal carcinoma HONE1 cell lysate was digested with MNase for release of the nucleosomes, and the histone-DNA complexes were immunoprecipitated with specific antibodies. The protein component in the precipitate was digested with proteinase K followed by DNA purification; the DNA library was constructed for sequence analysis.@*RESULTS@#Compared with the conventional DNA library construction, Tn5 transposase method allowed direct enrichment of the target DNA after Tn5 fragmentation, which was simple, time-saving and more efficient. The IGV visualized map showed that the information obtained by the two library construction methods was consistent. The sequencing data obtained by the two methods revealed more signal enrichment with Tn5 transposase library construction than with the conventional approach. H3K4me3 ChIP results showed a good reproducibility after Tn5 transposase library construction with a signal-to-noise ratio above 50%.@*CONCLUSIONS@#Tn5 transposase method improves the efficiency of DNA library construction and the results of subsequent sequence analysis, and is especially suitable for detecting histone modification in the DNA to provide a better technical option for epigenetic studies.
Subject(s)
Humans , Chromatin Immunoprecipitation , DNA , Gene Library , High-Throughput Nucleotide Sequencing , Reproducibility of Results , Sequence Analysis, DNAABSTRACT
Public library, academic library and special library are different kinds of library. The aim, objects, contents of service and the different characteristics, needs, problems of smart construction depend on their different functional positions. The problems and challenges faced by smart construction were pointed out based on the analy-sis of the origin, concept and characteristics of smart construction, domestic studies on smart construction, in com-bination with the different functional positions of public library, academic library and special library, and the needs of their smart construction. Suggestions were thus put forward for their solution.
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After the feasibility of hackerspace in medical library and its characteristics with teachers, medical staff and researchers as its main service objects, knowledge communication and sharing as its main service contents were analyzed, its implementation strategies in medical library were put forward, including definite positioning, rational planning, spreading and popularizing, and fund raising.
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As a new model of future library, the intelligent technology-supported smart library can be located at anywhere and at anytime. The current studies on domestic smart library were analyzed from its emerging background in order to provide reference for the relevant studies on smart library in our country.
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After the relationship between academic library and construction of think tank was analyzed, the role of academic library in construction of think tank was elaborated from the aspects of professionals, literature resources, operation mechanisms and cooperative development.
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After a description of ISO11620:2014 , the international criteria for library performance assessment , how to enforce the construction of military academic libraries was systematically elaborated in terms of improving the quality and use of their resources and infrastructures, and reducing their construction cost.
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After a description of the association of learning sciences with library and information science and the necessity to introduce learning sciences into library and information science , the role and profound influence of learning sciences in the development of modern library were summarized .
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Humanities education is an important part in college and university education. Medical college and university libraries should strengthen their sense of responsibility and sense of mission, continuously expand and innovate the mode of humanity education, improve the environment of teaching, modify the system construction and promote the quality of straits to make library as an important field for medical students humanity education.
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The paper focuses on how to establish a harmonious library based on people -oriented theory from two aspects: people -oriented reader service and librarian management, particularly including strengthening literature resources construction, reader service innovation, career planning and continuing education to librarians, etc.