1.Structure and function of the genome of coxsackievirus B3.
Wen-Qi HE ; Hui-Jun LU ; Feng GAO
Chinese Journal of Virology 2009;25(5):395-400
2.Detection of promoter and 3' UTR mutation in A20 gene of a case with T cell lymphoma cell leukemia.
Ling Ling ZHOU ; Geng Xin LUO ; Li Hua ZHU ; Qi WEI ; Yong Qiang WEI ; Ru FENG ; Yang Qiu LI
Chinese Journal of Hematology 2018;39(10):851-854
Objective: To clarify the characteristics of the A20 regulatory changes by analyzing mutations in the non-coding region of the A20 gene in patients with T-cell lymphoma leukemia (T-LCL) . Methods: PCR and nucleotide sequence analysis were used to detect mutations in the non-coding region of the A20 gene, and DNA samples from PBMCs of 52 cases of T-LCL and 99 healthy controls. Results: A missense mutation (c.-672T>G) was detected in the A20 gene promoter from one T-LCL patient, which has been registered as a SNP (rs139054966) in gene bank. Meanwhile, a new mutation was detected in the 3' UTR mRNA (3916 (C>G) ) . These two mutations were absent in other T-LCL samples and controls. Conclusion: The rs139054966 (c.-672T>G) and 3916 (C>G) mutations in the A20 gene were detected in T-LCL patients for the first time. There was also rs139054966 located on the binding region of the transcription factor P53, and its significance remained to be further clarified.
3' Untranslated Regions
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Humans
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Leukemia
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Lymphoma, T-Cell
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Mutation
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Promoter Regions, Genetic
3.Prediction of Mammalian MicroRNA Targets : Comparative Genomics Approach with Longer 3' UTR Databases.
Seungyoon NAM ; Young Kook KIM ; Pora KIM ; V Narry KIM ; Seokmin SHIN ; Sanghyuk LEE
Genomics & Informatics 2005;3(3):53-62
MicroRNAs play an important role in regulating gene expression, but their target identification is a difficult task due to their short length and imperfect complementarity. Burge and coworkers developed a program called TargetScan that allowed imperfect complementarity and established a procedure favoring targets with multiple binding sites conserved in multiple organisms. We improved their algorithm in two major aspects - (i) using well-defined UTR (untranslated region) database, (ii) examining the extent of conservation inside the 3' UTR specifically. Average length in our UTR database, based on the ECgene annotation, is more than twice longer than the Ensembl. Then, TargetScan was used to identify putative binding sites. The extent of conservation varies significantly inside the 3' UTR. We used the "tight" tracks in the UCSC genome browser to select the conserved binding sites in multiple species. By combining the longer 3' UTR data, TargetScan, and tightly conserved blocks of genomic DNA, we identified 107 putative target genes with multiple binding sites conserved in multiple species, of which 85 putative targets are novel.
3' Untranslated Regions*
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Binding Sites
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DNA
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Gene Expression
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Genome
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Genomics*
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Methods
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MicroRNAs*
4.Genetic Effect Analysis of β-globin Gene 3'UTR+101G>C (HBB:c. *233G>C) Variant.
Li DU ; Cui-Ze YAO ; Xiu-Qin BAO ; Jie LIANG ; Teng-Long YUAN ; Dan-Qing QIN ; Ji-Cheng WANG
Journal of Experimental Hematology 2021;29(4):1271-1274
OBJECTIVE:
To investigate whether β-globin gene 3'UTR+101G>C (HBB:c.*233G>C) variant has genetic effect and provide basis for gene diagnosis and genetic counseling.
METHOD:
Whole blood cell analysis and capillary zone electrophoresis (CZE) were used to analyze the hematological indexes. The most frequent 23 mutations in southern Chinese individuals were routinely measured by PCR-flow fluorenscence immunmicrobeads assay. Sanger sequencing was used to detect the other variants of β-globin gene (HBB).
RESULTS:
In 463 cases, a total of 7 cases with HBB:c.*233G>C variant were detected, among them 4 cases carried other pathogenic variants of HBB gene (2 cases were in trans, 2 cases were in cis), who had typical hematological characteristics of mild β-thalassemia, and 3 cases also carried abnormal hemoglobin variation, but did not have hematological characteristics of β-thalassemia.
CONCLUSION
The study shows that HBB:c.*233G > C variant has no obvious genetic effect and should be a benign polymorphism.
3' Untranslated Regions
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Hemoglobins, Abnormal/genetics*
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Humans
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Mutation
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beta-Globins/genetics*
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beta-Thalassemia/genetics*
5.Detection of pathogenic variants in four patients with globozoospermia.
Zhenzhen TANG ; Qingqin LI ; Guoyong CHEN ; Wujian HUANG ; Yulin WANG ; Yu YE ; Peng XIE ; Fenghua LAN ; Duo ZHANG
Chinese Journal of Medical Genetics 2023;40(3):301-307
OBJECTIVE:
To explore the genetic basis for 4 patients with globozoospermia.
METHODS:
Semen and blood samples were collected from the patients for the determination of sperm concentration, viability, survival rate, morphology and acrosome antigen CD46. Meanwhile, DNA was extracted for whole exome sequencing (WES), and candidate variants were validated by Sanger sequencing.
RESULTS:
All of the four patients were found to harbor variants of the DPY19L2 gene. Patients 1 ~ 3 had homozygous deletions of the DPY19L2 gene. Sanger sequencing confirmed that the DPY19L2 gene in patient 3 was disrupted at a recombination breakpoint area BP2, resulting in nonallelic homologous recombination and complete deletion of the DPY19L2 gene. Patients 2 and 3 respectively harbored novel homozygous deletions of exons 2 ~ 22 and exons 14 ~ 15. Patient 4 harbored heterozygous deletion of the DPY19L2 gene, in addition with a rare homozygous deletion of the 3' UTR region.
CONCLUSION
DPY19L2 gene variants probably underlay the globozoospermia in the four patients, which has fit an autosomal recessive pattern of inheritance and the characteristics of genomic diseases.
Male
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Humans
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Teratozoospermia/genetics*
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Homozygote
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Semen
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Sequence Deletion
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3' Untranslated Regions
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Membrane Proteins
6.MiR-4484 regulates the expression of integrin α 6 in gastric cancer tissues and its significance.
Hui Yu ZHONG ; Yuan ZHONG ; Yang WEN ; Xin Tong TAO ; Xing Bo SONG ; Xiao Jun LU
Chinese Journal of Oncology 2022;44(3):246-251
Objective: To investigate the relationship between the expression of integrin α 6 (ITGA6), miR-4484 and the pathologic stage of gastric cancer. Methods: Gastric cancer tissues and normal gastric mucosa tissues adjacent to cancer (>5 cm from tumor margin) of 30 patients with primary gastric cancer who underwent direct surgical resection without adjuvant therapy from June to September 2017 in West China Hospital of Sichuan University were selected. Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression levels of miR-4484 and ITGA6, western blot was used to detect the expression level of ITGA6 protein, dual luciferase reporter gene was used to verify the relationship between ITGA6 and miR-4484. Spearman's correlation analysis was used to determine the relationship between miR-4484 and ITGA6 expression levels in gastric cancer tissues. Results: The expression level of ITGΑ6 in gastric cancer (32.30±13.47) was higher than that in matched normal gastric tissues (24.55±10.25, P=0.015), the area under the receiver operating characteristic (ROC) curve was 0.660 and the diagnostic sensitivity and specificity were 43.3% and 96.7%, respectively. The expression level of miR-4484 in gastric cancer (4.11±2.87) was lower than that of matched normal gastric tissues (5.75±2.80, P=0.029), the area under the ROC curve was 0.690 and the diagnostic sensitivity and specificity were 30.0% and 86.7%, respectively. The expression level of miR-4484 was negatively correlated with ITGA6 in gastric cancer tissues (r=-0.621, P<0.001). The expression level of ITGA6 protein in gastric cancer tissues (0.65±0.19) was higher than that in normal adjacent tissues (0.26±0.12, P<0.001). Compared with ITGA6 3'UTR wild-type+ miR-NC group, ITGA6 3'UTR wild-type+ miRNA mimics group had lower luciferase activity (50.69±5.10, 34.00±1.19, P<0.001), while the luciferase activity of ITGA6 3'UTR wild-type+ ASO miR-4484 group was higher than that of ITGA6 3'UTR wild-type+ miR-NC group (82.44±6.37, 50.69±5.10, P<0.001), indicated that ITGA6 was the direct target gene of miR-4484. The expression levels of miR-4484 in T1, T2, T3 and T4 (4a and 4b) gastric cancer tissues were 9.98±2.24, 5.28±2.03, 2.92±2.04 and 4.11±2.87, respectively, with statistical significance (P<0.001). The expression levels of ITGA6 in N0, N1, N2 and N3 gastric cancer tissues were 29.55±8.32, 21.71±3.75, 24.60±8.79 and 40.69±15.83, respectively, with statistical significance (P=0.022). The expression levels of miR-4484 in N0, N1, N2 and N3 gastric cancer tissues were 5.01±3.52, 5.48±2.76, 5.88±1.83 and 2.30±1.56, respectively, with statistical significance (P=0.032). The expression levels of ITGA6 in M0 and M1 gastric cancer tissues were 26.28±7.66 and 52.08±8.12, respectively, with statistical significance (P<0.001). The expression levels of miR-4484 in M0 and M1 gastric cancer tissues were 4.95±2.74 and 1.34±0.80, respectively, with statistical significance (P<0.001). Conclusions: ITGA6 is upregulated in gastric cancer tissues, while miR-4484 is downregulated in the gastric cancer group, and its expression level is related to the clinicopathological features of gastric cancer. ITGA6 is the direct target gene of miR-4484, implicates that miR-4484 may inhibit the invasion and metastasis of gastric cancer by regulating the expression of ITGA6. Both miR-4484 and ITGA6 may be the new prognostic markers and potential therapeutic targets of gastric cancer.
3' Untranslated Regions
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China
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Humans
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Integrin alpha6/genetics*
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MicroRNAs/genetics*
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Stomach Neoplasms/pathology*
7.TT Virus Detection Using Different PCR Primer Sets in Healthy and Infected Individuals with Hepatitis B or C Viruses.
Han Sung KIM ; Jae Seok KIM ; Wonkeun SONG ; Hee Jung KANG ; Kyu Man LEE
Korean Journal of Clinical Microbiology 2007;10(1):14-18
BACKGROUND: TT virus (TTV) infection is highly prevalent in the general population and in the patients infected with hepatitis B virus (HBV) or hepatitis C vius (HCV). The aim of the present study was to assess the positive rates of TTV DNA using different PCR primer sets in healthy and HBV or HCV-infected individuals in Korea. METHODS: TTV DNA was investigated in serum samples of 69 healthy individuals and 59 HBV-infected and 34 HCV-infected individuals by nested PCR assays using primers from N22 region, 5'-untranslated region (UTR), and 3' UTR of viral genome. RESULTS: TTV DNA was detected in 43% of total study populations using N22 primers, in 69% using 5' UTR primers and, in 64% using 3' UTR primers. No significant difference was observed in the positive rates of TTV DNA between healthy and HBV or HCV- infected individuals. CONCLUSION: The PCR assays for TTV DNA using 5' UTR primers and 3' UTR primers exhibited higher positive rates than that of the assay using N22 primers without any significant difference between healthy and HBV or HCV-infected individuals.
3' Untranslated Regions
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5' Untranslated Regions
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DNA
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Genome, Viral
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Hepatitis B virus
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Hepatitis B*
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Hepatitis C
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Hepatitis*
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Humans
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Korea
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Polymerase Chain Reaction*
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Torque teno virus*
8.Structure and Expression Analyses of SVA Elements in Relation to Functional Genes.
Yun Jeong KWON ; Yuri CHOI ; Jungwoo EO ; Yu Na NOH ; Jeong An GIM ; Yi Deun JUNG ; Ja Rang LEE ; Heui Soo KIM
Genomics & Informatics 2013;11(3):142-148
SINE-VNTR-Alu (SVA) elements are present in hominoid primates and are divided into 6 subfamilies (SVA-A to SVA-F) and active in the human population. Using a bioinformatic tool, 22 SVA element-associated genes are identified in the human genome. In an analysis of genomic structure, SVA elements are detected in the 5' untranslated region (UTR) of HGSNAT (SVA-B), MRGPRX3 (SVA-D), HYAL1 (SVA-F), TCHH (SVA-F), and ATXN2L (SVA-F) genes, while some elements are observed in the 3'UTR of SPICE1 (SVA-B), TDRKH (SVA-C), GOSR1 (SVA-D), BBS5 (SVA-D), NEK5 (SVA-D), ABHD2 (SVA-F), C1QTNF7 (SVA-F), ORC6L (SVA-F), TMEM69 (SVA-F), and CCDC137 (SVA-F) genes. They could contribute to exon extension or supplying poly A signals. LEPR (SVA-C), ALOX5 (SVA-D), PDS5B (SVA-D), and ABCA10 (SVA-F) genes also showed alternative transcripts by SVA exonization events. Dominant expression of HYAL1_SVA appeared in lung tissues, while HYAL1_noSVA showed ubiquitous expression in various human tissues. Expression of both transcripts (TDRKH_SVA and TDRKH_noSVA) of the TDRKH gene appeared to be ubiquitous. Taken together, these data suggest that SVA elements cause transcript isoforms that contribute to modulation of gene regulation in various human tissues.
3' Untranslated Regions
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5' Untranslated Regions
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Exons
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Gene Expression Profiling
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Genome, Human
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Genomics
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Humans
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Lung
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Organ Specificity
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Poly A
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Primates
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Protein Isoforms
9.Molecular cloning and expression analysis of sucrose synthase gene from Dendrobium officinale.
Hengling MENG ; Chengli DUAN ; Fenghui XIAO ; Shengchao YANG ; Yinghong ZHA ; Guosong WEN
China Journal of Chinese Materia Medica 2011;36(7):833-837
OBJECTIVEClone of sucrose synthase of Dendribium officinale and expression analysis, to provide the theory basis for research the relationship between polysaccharide synthesis of D. officinale and sucrose synthase activity.
METHODAccording to homologous sequence of sucrose synthase gene on GenBank, application the technology of RT-PCR and RACE, clone the full length of D. officinale. Target gene amplified with T vector was transformed into competent E. coli. BL21, IPTG induced expression, SDS-PAGE analysis.
RESULTA full length cDNA encoding sucrose synthase was isolated from the D. officinale, named DOSS1, the GenBank accession number is HQ856835, the cDNA is 2781 bp in length containing an open reading frame of 2424 bp encoding 807 amino acids with a predicted molecular mass of 92.3 x 10(3), the deduced amino acid sequence of D. officinale sucrose synthase shares 95% identity with Mokara yellow (AF530568); shares 90% identity with Oncidium goldiana (AF530567); shares more than 80% with other monocotyledonous plants.
CONCLUSIONCloned the sucrose synthase gene and induced an obvious band successfully.
3' Untranslated Regions ; genetics ; 5' Untranslated Regions ; genetics ; Cloning, Molecular ; Dendrobium ; enzymology ; genetics ; metabolism ; Escherichia coli ; genetics ; Gene Expression Regulation, Plant ; Glucosyltransferases ; genetics ; metabolism ; Phylogeny ; Polysaccharides ; biosynthesis
10.The Role of MicroRNAs in Regulatory T Cells and in the Immune Response.
Immune Network 2011;11(1):11-41
The discovery of microRNA (miRNA) is one of the major scientific breakthroughs in recent years and has revolutionized current cell biology and medical science. miRNAs are small (19~25nt) noncoding RNA molecules that post-transcriptionally regulate gene expression by targeting the 3' untranslated region (3'UTR) of specific messenger RNAs (mRNAs) for degradation of translation repression. Genetic ablation of the miRNA machinery, as well as loss or degradation of certain individual miRNAs, severely compromises immune development and response, and can lead to immune disorders. Several sophisticated regulatory mechanisms are used to maintain immune homeostasis. Regulatory T (Treg) cells are essential for maintaining peripheral tolerance, preventing autoimmune diseases and limiting chronic inflammatory diseases. Recent publications have provided compelling evidence that miRNAs are highly expressed in Treg cells, that the expression of Foxp3 is controlled by miRNAs and that a range of miRNAs are involved in the regulation of immunity. A large number of studies have reported links between alterations of miRNA homeostasis and pathological conditions such as cancer, cardiovascular disease and diabetes, as well as psychiatric and neurological diseases. Although it is still unclear how miRNA controls Treg cell development and function, recent studies certainly indicate that this topic will be the subject of further research. The specific circulating miRNA species may also be useful for the diagnosis, classification, prognosis of diseases and prediction of the therapeutic response. An explosive literature has focussed on the role of miRNA. In this review, I briefly summarize the current studies about the role of miRNAs in Treg cells and in the regulation of the innate and adaptive immune response. I also review the explosive current studies about clinical application of miRNA.
3' Untranslated Regions
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Adaptive Immunity
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Autoimmune Diseases
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Cardiovascular Diseases
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Gene Expression
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Homeostasis
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Immune System Diseases
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MicroRNAs
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Peripheral Tolerance
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Prognosis
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Repression, Psychology
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RNA, Messenger
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RNA, Untranslated
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T-Lymphocytes, Regulatory