1.Stable study of gene sequence of seed lot system G1P4 (2001019210) during production of rotavirus vaccine
Luan Thi Le ; Hien Dang Nguyen ; Huong Thi Mai Nguyen
Journal of Preventive Medicine 2008;18(6):5-11
Background: Presently, toxicity decreased oral live rotavirus is a candidate for vaccine for the prevention of rotavirus induced diarrhea. According to the World Health Organization, the seed lot system is robustly checked, in which determining the stable of gene sequence. Objective: To determine the sequence of genes 4: 6: 9: 10 with base pair correlative 855: 824: 1314: 734 of seed lot system G1P4 (2001019210) during production of rotavirus vaccine. Subject and methods: Gene 4 (VP4), gene 6 (VP6), gene 9 (VP7) and gene 10 (NSP4) of seed lot system G1P4 were determined for gene sequencing and then comparing the nucleotide sequence as well as deduced amino acids from original strain with the produced strain and vaccine virus. Results and Conclusion: There was no different for nucleotide and deduced amino acid sequence from the original strain during the production of rotavirus vaccine of G1P4 MS (2001019210) to producing strains of G1P4 WS and vaccine strains of G1P4 VX.
rotavirus vaccine
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gene sequence
2.Study on the stable of gene sequence of seed lot system G1P8 KH0118 during production rotavirus vaccine
Luan Thi Le ; Hien Dang Nguyen ; Huong Thi Mai Nguyen
Journal of Preventive Medicine 2008;18(6):67-73
Background: Rotavirus strain (KH0118) is used as the primary material to produce original rotavirus vaccine strains with the symbol of G1P8 MS. According to the World Health Organization\u2019s standard, the strain is needed to evaluate the stability of gene throughout analysis of gene and amino acid sequence during vaccine production. Objective: To determine the sequence of genes 4 (VP4), 6 (VP6), 9 (VP7) and 10 (NSP4) with base pair correlative 855:866:1345:745 of seed lot system and vaccine of G1P8 strain and to evaluate the stability of seed lot system during vaccine production. Subject and methods: ARN was divided from the original strain of rotavirus vaccine G1P8 MS, rotavirus vaccine productive strain (G1P8 WS) and rotavirus vaccine (G1P8 VX). Then using primer pairs to determine gene sequence VP4, VP6, VP7, NSP4 and comparing gene and amino acid sequence of the seed lot system. Results and Conclusion: The study demonstrated that, there was no difference for the nucleotide and amino acid sequence from the original strain during production of rotavirus vaccine G1P8 KH0118.
rotavirus vaccine
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G1P8
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gene sequence
3.HisCoM-PAGE: software for hierarchical structural component models for pathway analysis of gene expression data
Genomics & Informatics 2019;17(4):45-
To identify pathways associated with survival phenotypes using gene expression data, we recently proposed the hierarchical structural component model for pathway analysis of gene expression data (HisCoM-PAGE) method. The HisCoM-PAGE software can consider hierarchical structural relationships between genes and pathways and analyze multiple pathways simultaneously. It can be applied to various types of gene expression data, such as microarray data or RNA sequencing data. We expect that the HisCoM-PAGE software will make our method more easily accessible to researchers who want to perform pathway analysis for survival times.
Gene Expression
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Methods
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Phenotype
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Sequence Analysis, RNA
4.Development and verification of an FLP/FRT system for gene editing in Bacillus licheniformis.
Zongwen LI ; Youran LI ; Zhenghua GU ; Zhongyang DING ; Liang ZHANG ; Sha XU ; Guiyang SHI
Chinese Journal of Biotechnology 2019;35(3):458-471
Few tools of gene editing have been developed in Bacillus licheniformis at present. In order to enrich the tools, an FLP/FRT gene editing system that can repeatedly use a single selectable marker was constructed in Bacillus licheniformis, and the system was verified by knocking out an alpha amylase gene (amyL), an protease gene (aprE) and knocking in an exogenous Vitreoscilla hemoglobin gene (vgb). First, knock-out plasmids pNZTT-AFKF of amyL and pNZTT-EFKF of aprE were constructed using thermosensitive plasmid pNZT1 as a carrier. The two knock-out plasmids contained respective homology arms, resistance genes and FRT sites. Then the knock-out plasmids were transformed into Bacillus licheniformis and the target genes were replaced by respective deletion cassette via twice homologous exchange. Finally, an expression plasmid containing FLP recombinase reading frane was introduced and mediated the excision of resistance marker. In order to expand the practicability of the system, knock-in plasmid pNZTK-PFTF-vgb was constructed, with which knock-in of vgb at pflB site was carried out successfully. The results showed that amyL and aprE were successfully knocked out and the marker kanamycin cassette exactly excised. The activities of amylase and protease of deletion mutants were reduced by 95.3% and 80.4% respectively. vgb was successfully knocked in at pflB site and the marker tetracycline cassette excised. The expression of integrated vgb was verified via real-time PCR. It is the first time to construct an FLP/FRT system for gene editing in Bacillus licheniformis, which could provide an effective technical means for genetic modification.
Bacillus licheniformis
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Gene Editing
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Plasmids
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Sequence Deletion
5.A review on integration methods for single-cell data.
Duo PAN ; Huamei LI ; Hongde LIU ; Xiao SUN
Journal of Biomedical Engineering 2021;38(5):1010-1017
The emergence of single-cell sequencing technology enables people to observe cells with unprecedented precision. However, it is difficult to capture the information on all cells and genes in one single-cell RNA sequencing (scRNA-seq) experiment. Single-cell data of a single modality cannot explain cell state and system changes in detail. The integrative analysis of single-cell data aims to address these two types of problems. Integrating multiple scRNA-seq data can collect complete cell types and provide a powerful boost for the construction of cell atlases. Integrating single-cell multimodal data can be used to study the causal relationship and gene regulation mechanism across modalities. The development and application of data integration methods helps fully explore the richness and relevance of single-cell data and discover meaningful biological changes. Based on this, this article reviews the basic principles, methods and applications of multiple scRNA-seq data integration and single-cell multimodal data integration. Moreover, the advantages and disadvantages of existing methods are discussed. Finally, the future development is prospected.
Base Sequence
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Gene Expression Profiling
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Gene Expression Regulation
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Humans
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Sequence Analysis, RNA
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Single-Cell Analysis
6.Cloning of full length cDNA sequence of the mouse ameloblastin.
Shuping GU ; Junnan SHI ; Han LIU ; Jianjun HAO ; Ping WANG ; Jian FEI
Chinese Journal of Stomatology 2002;37(4):278-280
OBJECTIVEScreening for special genes of matrix proteins of dentin and enamel of mouse dental germ.
METHODSA cDNA library of dental germ of mouse was screened by differential display. The interesting clones were sequenced.
RESULTSSix positive clones were isolated from the cDNA library. The sequence of one of the six positive clones was homologous with the ameloblastin sequence of rat. There are 497 homologous base pairs between the 526 base pairs sequenced by pTriplEX 3' primer of this clone and the 32-580 sequence of the rat ameloblastin gene; and there are 533 homologous base pairs between the 567 base pairs sequenced by pTriplEX 5' primer of this clone and the 1285-1854 sequence of the rat ameloblastin gene.
CONCLUSIONSThe full length cDNA sequence of the mouse ameloblastin was cloned.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Gene Library ; Mice ; Molecular Sequence Data
7.TRAPR: R Package for Statistical Analysis and Visualization of RNA-Seq Data.
Jae Hyun LIM ; Soo Youn LEE ; Ju Han KIM
Genomics & Informatics 2017;15(1):51-53
High-throughput transcriptome sequencing, also known as RNA sequencing (RNA-Seq), is a standard technology for measuring gene expression with unprecedented accuracy. Numerous bioconductor packages have been developed for the statistical analysis of RNA-Seq data. However, these tools focus on specific aspects of the data analysis pipeline, and are difficult to appropriately integrate with one another due to their disparate data structures and processing methods. They also lack visualization methods to confirm the integrity of the data and the process. In this paper, we propose an R-based RNA-Seq analysis pipeline called TRAPR, an integrated tool that facilitates the statistical analysis and visualization of RNA-Seq expression data. TRAPR provides various functions for data management, the filtering of low-quality data, normalization, transformation, statistical analysis, data visualization, and result visualization that allow researchers to build customized analysis pipelines.
Base Sequence
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Gene Expression
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Gene Expression Profiling
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Molecular Sequence Data
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Programming Languages
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Sequence Analysis, RNA
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Statistics as Topic
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Transcriptome
8.Global Optimization of Clusters in Gene Expression Data of DNA Microarrays by Deterministic Annealing.
Kwon Moo LEE ; Tae Su CHUNG ; Ju Han KIM
Genomics & Informatics 2003;1(1):20-24
The analysis of DNA microarry data is one of the most important things for functional genomics research. The matrix representation of microarray data and its successive 'optimal' incisional hyperplanes is a useful platform for developing optimization algorithms to determine the optimal partitioning of pairwise proximity matrix representing completely connected and weighted graph. We developed Deterministic Annealing (DA) approach to determine the successive optimal binary partitioning. DA algorithm demonstrated good performance with the ability to find the 'globally optimal' binary partitions. In addition, the objects that have not been clustered at small non-zero temperature, are considered to be very sensitive to even small randomness, and can be used to estimate the reliability of the clustering.
Cluster Analysis
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DNA*
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Gene Expression*
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Genomics
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Oligonucleotide Array Sequence Analysis*
10.Serial analysis of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues.
Yao-fei HU ; Zu-yuan REN ; Yun-feng LI ; Hong-xia SUN ; Yong-sheng CHANG ; Chang-bao SU ; Ren-zhi WANG ; Jin ZUO ; Fu-de FANG
Acta Academiae Medicinae Sinicae 2002;24(6):611-615
OBJECTIVETo observe the characteristics and difference of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues.
METHODSUsing serial analysis of gene expression (SAGE), two SAGE libraries were generated. Forty clones from each SAGE library were sequenced, and the results were analyzed by SAGE2000 software and compared with the SAGE map at NCBI.
RESULTSA total of 655 gene tags, representing 43 genes, were extracted from the 40 sequence files of the para-tumor normal pituitary tissues and 737 gene tags, representing 53 genes, were extracted from the 40 sequence files of the pituitary adenomas. Of these tags, 13 were not reported before. The genes related to pituitary hormone secretion and energy metabolism were highly expressed in the two kinds of tissues. Some growth factors and cytokines were also expressed, including those involved in the immunological system. But there were also much difference of gene expression in the two tissues. Thirty-one and five tags were only detected in para-tumor normal pituitary tissues and pituitary adenomas, respectively.
CONCLUSIONSGenes involved in hormones secretion and energy metabolism were highly expressed in the pituitary adenomas and para-tumor normal pituitary tissues. Many growth factors and cytokines were also expressed in pituitary. There was also much difference of gene expression in the two kinds of tissues. SAGE can be used not only in understanding the quantity information of gene expression, but also in finding new genes.
Adenoma ; genetics ; metabolism ; Base Sequence ; Cloning, Molecular ; Expressed Sequence Tags ; Gene Expression ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Pituitary Gland ; metabolism ; Pituitary Neoplasms ; genetics ; metabolism