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Mutations and intron polymorphisms in voltage-gated sodium channel genes of different geographic populations of Culex pipiens pallens/ Culex pipiens quinquefasciatus in China

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贫困所致传染病(英文) 2024年第2期13卷 6-16页

作者： Li Wenyu Ma Delong Mu Qunzheng Zhou Xinxin Hua Dongdong Zhao Chunchun Liu Qiyong Wang Jun Meng Fengxia State Key Laboratory of Infectious Disease Prevention and Control Department of Vector Biology and Control National Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and Prevention WHO Collaborating Centre for Vector Surveillance and Management Beijing People’s Republic of China Ningxia Center for Disease Prevention and Control YinChuan NingXia People’s Republic of China Jinan Shizhong Center for Disease Control and Prevention Jinan Shandong China Vanke School of Public Health Tsinghua University Beijing People’s Republic of China Weifang No. People’s Hospital Weifang Shandong People’s Republic of China Beijing Daxing Center for Disease Control and Prevention Beijing People’s Republic of China Jinan Second Maternal and Child Health Hospital Jinan Shandong China

Background Culex pipiens pallens andCulex pipiens quinquefasciatus are the dominant species ofCulex mosquitoes in China and important disease vectors. Long-term use of insecticides can cause mutations in the voltage-gated sodium channel (vgsc) gene of mosquitoes, but little is known about the current status and evolutionary origins ofvgsc gene in different geographic populations. Therefore, this study aimed to determine the current status ofvgsc genes inCx. p. pallens andCx. p. quinquefasciatus in China and to investigate the evolutionary inheritance of neighboring downstream introns of thevgsc gene to determine the impact of insecticides on long-term *** Sampling was conducted from July to September 2021 in representative habitats of 22 provincial-level administrative divisions in China. Genomic DNA was extracted from 1308 mosquitoes, the IIS6 fragment of thevgsc gene on the nerve cell membrane was amplified using polymerase chain reaction, and the sequence was used to evaluate allele frequency and knockdown resistance (kdr) frequency. MEGA 11 was used to construct neighborjoining (NJ) tree. PopART was used to build a TCS *** There were 6 alleles and 6 genotypes at the L1014 locus, which included the wild-type alleles TTA/L and CTA/L and the mutant alleles TTT/F, TTC/F, TCT/S and TCA/S. The geographic populations with akdr frequency less than 20.00% were mainly concentrated in the regions north of 38° N, and the geographic populations with akdr frequency greater than 80.00% were concentrated in the regions south of 30° *** frequency increased with decreasing latitude. And within the same latitude, the frequency ofkdr in large cities is relatively high. Mutations were correlated with the number of introns. The mutant allele TCA/S has only one intron, the mutant allele TTT/F has three introns, and the wild-type allele TTA/L has 17 *** Cx. p. pallens andCx. p. quinquefasciatus have developed resistance to insecticides in mos

关键词： Culex pipiens pallens Culex pipiens quinquefasciatus vgsc gene Mutation Frequency intron Haplotype China

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Analysis of intron sequence variability of the conservative HMG-box of Sox9 genes in allotetraploids and their original parents

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Progress in Natural Science:Materials International 2007年第5期17卷 537-543页

作者： Liu Jifang, Liu Shaojun , Tao Min, Li Wei and Liu Yun (Key Laboratory of Protein Chemistry and Developmental Biology of State Education Ministry of China, College of Life Sciences, Hunan Normal University, Changsha 410081, China) Key Lab. of Protein Chemistry and Developmental Biology MOE College of Life Sciences Hunan Normal University Changsha 410081 China

The Sox genes of allotetraploids and their original maternal red crucian carp (Carassius caassius red var.) and original paternal common carp (Cyprinus carpio L.) were detected by PCR with the designed primers based on the conserved HMG-box sequence in different species. Sequencing of Sox genes indicated that two Sox9 genes (Atsox9a and Atsox9b) existed in allotetraploids, while only one Sox9 gene existed in red crucian carp (Rcsox9a) and common carp (Ccsox9b) . All of the four Sox9 genes contained an intron in the HMG-box. with the sizes of 413 bp, 703 bp, 401 bp and 714 bp, respectively. Moreover, the introns obeyed the rule of "GT-AG" . A high similarity was observed between introns of Atsox9a and Rcsox9a (94.4%), Atsox9b and Ccsox9b (97.8%). Interestingly, the deduced amino acid sequences of their corresponding exons all shared 100% identity. Thus, introns of the HMG-domain of Sox9s in allotetraploids and their original parents have not only the length polymorphism but also intron variability. Our results provide significant molecular evidence for the origin and evolution of allotetraploids.

关键词： Sox9 intron genetic variability phylogenesis.

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Construction and expression of inverted configuration of retroviral vector containing intron 1 of hFIX

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Chinese Science Bulletin 1998年第4期43卷 315-318页

作者： Wang Hongwei Bao Yun Xin Yongna Yang Xiaoqin Shi Qian Lu Daru Qiu Xinfang Xue Jinglun Fudan Univ Inst Genet Shanghai 200433 Peoples R China

intron was found to play an important role in improving gene expression. To improve the human factor IX(hFIX) expression level in hemophilia B gene therapy study, the retroviral vector containing intron 1 of hFIX gene was constructed in forwarded configuration, but the intron 1 was found spliced in virus particles by RT_PCR detection. So the inverted configuration vector G1NaPAi′IX was suggested and constructed on the basis of SNMBAIXm and transfected into PA317. Then C2C12 cells were transfected using the above virus supernatant and the G418_resistant clones were selected. PCR and RT_PCR detection found that intron 1 structure existed in C2C12 clones and retroviral particles. And the expression level of inverted vector was 3 times higher than that of forwarded vector. These results showed that the inverted configuration vector was in deed able to avoid splicing of intron 1 during the process of retroviral packaging and improved the expression level of hFIX protein.

关键词： hFIX cDNA intron 1 retroviral vector gene therapy.

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Functional Characterization of the Promoter and Second intron of CUM1 During Flower Development in Cucumber(Cucumis sativus L.)

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Horticultural Plant Journal 2018年第3期4卷 103-110页

作者： GU Ran LIU Xiaofeng ZHAO Wensheng YAN Shuangshuang SUN Linhan WU Binning ZHANG Xiaolan Department of Vegetable Sciences Beijing Key Laboratory of Growth and Developmental Regulation for Protected Vegetable CropsChina Agricultural University

The characterization of flower specific promoter is critical during flower development by cucumber transformation ***(AG)is an organ identity gene that is required for carpel and stamen development in *** promoter and second intron of AG contain multiple regulatory elements that confer proper spatial and temporal *** is an important vegetable with unisexual *** MADS-box 1(CUM1)is the AG homolog in cucumber,belonging to the eu AG lineage along with *** situ hybridization showed that CUM1 was specifically expressed in the stamens and carpels of *** staining indicated that the second intron of CUM1 confers stamen-specific expression,while the promoter of CUM1 drives both stamen-and carpel-specific expression during the early stages of flower development,but is restricted to carpel-and connectivum-specific expression during the late stages of flower ***,a yeast one-hybrid assay demonstrated that two auxin response factors(Cs ARF13 and Cs ARF17)had bound directly to the second intron of *** data suggest that different regulatory circuits operate in AG homologs in plant species with distinct sex types.

关键词： cucumber CUM1 promoter intron in situ hybridization GUS staining yeast one-hybrid

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Isolation of tomato proteinase inhibitor Ⅱ gene and the function of its intron

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Chinese Science Bulletin 2002年第10期47卷 830-834页

作者： Xie, XZ Wu, NH Chinese Acad Sci Inst Dev Biol Beijing 100080 Peoples R China

The genomic DNA sequence of tomato pro-teinase inhibitor II gene (named tin2i, whose accession number in GenBank is AF007240) was isolated by PCR techniques. The intron sequence (TPI), with a length of 109 bp, owns typical structures of GT/AG dinucieotides at both ends and high content of AT base pairs which accounts for 80.7% of the total nucleotides. As shown by recombination experiment, the TPI sequence could efficiently promote the expression of the reporter gene gusA and this effect was independent of the position and orientation of the intron, thus showing its role as an enhancer. Such experiments as gel retardation assays, GUS histochemical staining and GUS fluorometric assays further demonstrated that TPI sequence maybe has promoter-like activity.

关键词： tomato proteinase inhibitor II gene intron enhancer.

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Phylogenetic relationships of Cyprinidae (Teleostei:Cypriniformes) inferred from the partial S6K1 gene sequences and implication of indel sites in intron 1

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Science China(Life Sciences) 2007年第6期50卷 780-788页

作者： KONG XiangHui1,2, WANG XuZhen1, GAN XiaoNi1, LI JunBing1 & HE ShunPing1 1 Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China 2 College of Life Science, Henan Normal University, Xinxiang 453007, China Institute of Hydrobiology Chinese Academy of Sciences Wuhan China College of Life Science Henan Normal University Xinxiang China

The family Cyprinidae is widely distributed in East Asia, and has the important phylogenetic signifi- cance in the fish evolution. In this study, the 5′ end partial sequences (containing exon 1, exon 2 and indel 1) of S6K1 gene were obtained from 30 representative species in Cyprinidae and outgroup using PCR amplification and sequencing. The phylogenetic relationships of Cyprinidae were reconstructed with neighbor joining (NJ), maximum parsimony (MP), maximum likelihood (ML), and Bayesian meth- ods. Myxocyprinus asiaticus (Catostomidae) was assigned to the outgroup taxon. Similar phylogenetic relationships within the family Cyprinidae were achieved with the four analyses. Leuciscini and Barbini were monophyletic lineages respectively with the high nodal supports. Leuciscini comprises Hy- pophthalmichthyinae, Xenocyprinae, Cultrinae, Gobioninae, Acheilognathinae and East Asian species of Leuciscinae and Danioninae. Monophyly of East Asian clade was supported with high nodal support. Barbini comprises Schizothoracinae, Barbinae, Cyprininae and Labeoninae. The monophyletic lineage consisting of Danio rerio, D. myersi, and Rasbora trilineata was basal in the tree. In addition, the large fragment indels in intron 1 were analyzed to improve the understanding of Cyprinidae relationships. The results showed that the large fragment indels were correlated with the relations among species. Some conserved regions in intron 1 were thought to be involved in the functional regulation. However, no correlation was found between sequence variations and species characteristic size.

关键词： Cyprinidae, S6K1 gene, phylogenetic relationships, intron

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First intron retention in part transcripts of OsEBP-89 gene in tissues of rice

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Chinese Science Bulletin 2002年第5期47卷 394-397页

作者： Huijun Yang Zongyang Wang Jingliu Zhang Xiuling Cai Mengmin Hong Acad Sinica Shanghai Inst Organ Chem State Key Lab Plant Mol Genet Shanghai 200032 Peoples R China

OsEBP-89 is a transcription factor gene of rice. It contains two introns. Using RT-PCR and Southern hybridization to study OsEBP-89 tissue-specific expression, we found that its first intron (115 bp in length) of its was retained in a fraction of its transcripts of this gene in rice developing seeds. Furthermore, two OsEBP-89 cDNA clones (c89L and c89LH) were screened from a rice cDNA library. Sequence analysis revealed that the first intron was retained in c89L clone, whereas, both the first and second intron sequences were spliced in c89LH. In addition to developing seeds, the first intron unspliced transcripts of OsEBP-89 are detected in leaves and roots of rice, too. However, the ratio of the first intron unspliced to spliced OsEBP-89 transcripts varied in different tissues examined. The potential biological significance of intron retention in OsEBP-89 transcript was discussed.

关键词： Oryza sativa transcription factor intron retention.

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Statistical characteristics of eukaryotic intron database

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Frontiers in Biology 2006年第4期1卷 362-366页

作者： HE Miao LI Jidong ZHANG Shanghong School of Life Sciences Sun Yat-Sen UniversityGuangzhou 510275China The Key Laboratory of Gene Engineering of Ministry of Education and Biotechnology Research CenterSun Yat-Sen UniversityGuangzhou 510275China

A database called eukaryotic intron database(EID)was developed based on the data from *** on the statistical characteristics of EID show that there were 103,848 genes,478,484 introns,and 582,332 exons,with an average of 4.61 introns and 5.61 exons per *** of 40−120 nt in length were abundant in the *** of the statistical analysis on the data from nine model species showed that in eukaryotes,higher species do not necessarily have more introns or exons in a gene than lower ***,characteristics of EID,such as intron phase,distribution of different splice sites,and the relationship between genome size and intron proportion or intron density,have been studied.

关键词： eukaryote intron database statistical characteristics

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The Sequence Variations of intron-3 of the α-Amylase Gene in Adzuki Bean

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Agricultural Sciences in China 2003年第10期2卷 1076-1082页

作者： JINWen-lin YamaguchiHirofumi IsigamiMatiko YasudaKentaro InstituteofCropGeneticBreeding BeijingAgriculturalCollegeBeijing102206P.R.China ConservationEcology GraduateSchoolofAgricultureandBiologicalSciencesOsakaPrefectureUniversitySakaiOsaka599-8531Japan

This study describes variation of intron-3 of α-amylase gene from 156 breeds of adzuki beans using SSCP(single-strand conformation polymorphism)analysis. Based on α-amylase gene structure and sequence, A pair of PCR primers, F (CCTACATTCTAACACACCCT) and R (GCATATTGTGCCAGTACAAT) were designed to amplify intron-3 fragments of α-amylase gene. 14 variant types were detected, including 13, 9, 10, 4 variant types in the wild, weed, locally cultivated and modern brought-up adzuki beans respectively, 9, 8, 7 variant types of the wild adzuki beans from Japan, China and Korea respectively, and some other variant types in the local adzuki beans from China and Bhutan. 60% of subjects of cultivated races were found to be EE type in the experiment. In addition, sequence analysis of intron-3 of α-amylase gene from 8 variant types reveals the evolution process of various variant types in adzuki beans.

关键词： Adzuki bean (Vigna angularis) SSCP (single-strand conformation polymorphism) α- Amylase gene intron Variation

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Constructing Competitive Reverse Transcription Polymerize Chain Reaction Inter-Reference of PC mRNA by intron Approach

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Agricultural Sciences in China 2004年第7期3卷 548-552页

作者： XUChuang XIACheng LIUGuo-wen WANGZhe JIANGYu-fu ZHANGNai-shen~ FUShi-xin FacultyofMilitaryVeterinaryMedicine QuartermasterUniversityofChinesePeople＇sLiberationArmyChangchun130062P.R.China

Inter-reference of competitive reverse transcription polymerase chain reaction (RT-PCR)was constructed by intron method to detect the change of PC mRNA level in the pathway ofcarbohydrate metabolism. The experiment based on the principle that 81bp intron sequencewas deleted in PC mRNA compared with PC DNA sequence. The 466bp competitive DNA templaterecombinant plasmid of PC mRNA was successfully built by a pair of primer and was clonedonce, PC DNA and PC mRNA could be inter-referred each other. The intron approach used inthe experiment has broken through the traditional method of constructing competitivetemplate.

关键词： PC Competitive template intron Quantitative RT-PCR Clone

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