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藏绵羊乳腺溶菌酶基因的克隆、原核表达及抗菌活性研究

藏绵羊乳腺溶菌酶基因的克隆、原核表达及抗菌活性研究

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：李建波江明锋王永青藏高原动物遗传资源保护与利用四川省重点实验室西南民族大学生命科学与技术学院

溶菌酶(LZM,EC 3.2.1.17)是富含半胱氨酸的抗菌蛋白之一,在哺乳动物非特异性免疫中起重要作用。本研究利用RT-PCR方法克隆了藏绵羊LZM基因cDNA序列,并成功构建原核表达质粒pET-32α/LZM,得到高效稳定表达的BL21菌株,抗菌实验表明重组LZ... 详细信息

溶菌酶(LZM,EC 3.2.1.17)是富含半胱氨酸的抗菌蛋白之一,在哺乳动物非特异性免疫中起重要作用。本研究利用RT-PCR方法克隆了藏绵羊LZM基因cDNA序列,并成功构建原核表达质粒pET-32α/LZM,得到高效稳定表达的BL21菌株,抗菌实验表明重组LZM有较强的抗金黄色葡萄球菌活性。系统进化树分析表明,南LZM氨基酸序列构建的物种进化关系与动物分类关系基本一致,其中藏绵羊乳腺LZM与山羊LZM的亲缘关系最近。实时荧光定量PCR(RT-qPCR)结果表明,藏绵羊乳腺LZM在乳腺中表达量最大,在气管、皱胃、小肠和肝中也有表达。为将来进一步研究LZM奠定了基础。

关键词：藏绵羊 LZM 克隆序列分析原核表达抗菌活性

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眼皮肤白化病基因突变研究及其在产前诊断中的应用

眼皮肤白化病基因突变研究及其在产前诊断中的应用

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：刘治佑吴东康冰侯巧芳姜锦成廖世秀河南省医学遗传研究所郑州大学人民医院河南省人民医院

白化病(albinism)是一组由黑素合成相关的基因突变导致眼或眼、皮肤、毛发黑色素缺乏引起的遗传性疾病。全身各部位均有色素缺乏的称为眼皮肤白化病(OCA),由于基因缺陷导致黑色素生物合成减少或完全缺乏,主要表现为眼、皮肤、毛发黑色... 详细信息

白化病(albinism)是一组由黑素合成相关的基因突变导致眼或眼、皮肤、毛发黑色素缺乏引起的遗传性疾病。全身各部位均有色素缺乏的称为眼皮肤白化病(OCA),由于基因缺陷导致黑色素生物合成减少或完全缺乏,主要表现为眼、皮肤、毛发黑色素缺乏和弱视等。患者全身皮肤呈白色或粉红色,毛发呈白色或淡黄色,具有视力低下、畏光、眼球震颤等表现。本文应用PCR技术扩增酪氨酸酶(TYR)基因外显子区域,DNA序列测序技术分析先证者及其父母的基因突变,确定致病性突变,并对再次怀孕的先证者母亲进行产前诊断。结果显示先证者TYR基因型为c.929 930insC,其父母分别为TYR基因c.929 930insC突变杂合了,符合常染色体隐形遗传规律。先证者母亲再次怀孕8周,待其孕20周后抽取羊水,对胎儿进行产前诊断。本研究对白化病相关基因诊断、产前基因诊断、携带行检测及遗传咨询等方面具有重要的意义。

关键词：白化病眼皮肤白化病基因突变产前诊断

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籼稻成熟种子组织培养特性的遗传分析和QTL定位

籼稻成熟种子组织培养特性的遗传分析和QTL定位

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：阮班普田福宽颜美仙叶世芳彭友林董国军朱丽胡江颜红岚郭龙彪钱前高振宇中国水稻研究所水稻生物学国家重点实验室

以籼稻品种杨稻6号和培矮64s为亲本的重组自交系,采用SSR、STS、CAPS和SNP共四种类型的分子标记构建了遗传图谱,覆盖水稻基因组1732.5 cM,标记问平均距离为12cM。用水稻成熟种子进行组织培养,对愈伤组织诱导率、褐化率和愈伤组织增重这... 详细信息

以籼稻品种杨稻6号和培矮64s为亲本的重组自交系,采用SSR、STS、CAPS和SNP共四种类型的分子标记构建了遗传图谱,覆盖水稻基因组1732.5 cM,标记问平均距离为12cM。用水稻成熟种子进行组织培养,对愈伤组织诱导率、褐化率和愈伤组织增重这三个指标进行了测定。QTL分析结果显示,在水稻第7和第10号染色体上检测出3个控制愈伤组织诱导率的QTL;在水稻第6、第7、第9号染色体上各检测到1个控制愈伤组织褐化率的QTL;在水稻第5和第7号染色体上分别检测到1个愈伤组织增重QTL。其中,水稻第7号染色体SSR标记RM5481和RM6835之间同时检测到愈伤组织诱导率QTL和褐化率QTL,分别解释7.29%和12.52%的表型变异。进一步的精细定位正在进行中。

关键词：籼稻组织培养 QTL定位

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Let-7参与DNA损伤应答的分子机理研究

Let-7参与DNA损伤应答的分子机理研究

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：谢晨蔡秋娴蒋嵩山中山大学生命科学学院有害生物控制与资源利用国家重点实验室中山大学生命科学学院基因工程教育部重点实验室

MicroRNAs(miRNAs)是一类小非编码RNA,广泛参与基因转录后水平调控。研究表明let-7 miRNA在肿瘤的发生发展中扮演重要角色,但其分子调控机理并不完全清楚。我们发现在神经胶质瘤细胞中,导致DNA损伤的试剂或化疗药物能够上调let-7的表达... 详细信息

MicroRNAs(miRNAs)是一类小非编码RNA,广泛参与基因转录后水平调控。研究表明let-7 miRNA在肿瘤的发生发展中扮演重要角色,但其分子调控机理并不完全清楚。我们发现在神经胶质瘤细胞中,导致DNA损伤的试剂或化疗药物能够上调let-7的表达水平和下调MDM4的表达水甲。靶基基因预测和细胞共转染试验表明let-7通过直接结合MDM4基因的编码区下调MDM4的表达,从而参与了调控p53信号转导分子通路。而此前的神经胶质瘤患者基因突变检测表明let-7在MDM4基因编码区上的作用位点存在突变,这暗示MDM4基因上的这个突变呵能会导致let-7失去对MDM4的调控能力,从而影响let-7在肿瘤发生发展中的作用。

关键词： let-7 DNA损伤 MDM4

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Molecular analysis of a deletion hotspot in the NRXN1 region reveals the involvement of short inverted repeats in deletion CNVs

Molecular analysis of a deletion hotspot in the NRXN1 region...

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：陈晓丽 Yiping Shen Feng Zhang Colby Chiang Vamsee Pillalamarri Ian Blumenthal Michael Talkowski Bai-Lin Wu James F.Gusella Capital Institute of Pediatrics Center for Human Genetic Research Massachusetts General Hospital Department of Laboratory Medicine Children's Hospital Boston Shanghai Children's Medical Center Shanghai Jiaotong University School of Medicine Department of Pathology Harvard Medical School State Key Laboratory of Genetic Engineering and Ministry of Education Key Laboratory of Contemporary Anthropology.School of Life Sciences Fudan University Department of Neurology Harvard Medical School Broad Institute of MIT and Harvard Children's Hospital and Institutes of Biomedical Science FudanUniversity Department of Genetics Harvard Medical School

NRXN1 microdeletions occur at relatively high frequency and confer increased risk for neurodevelopmental and neurobehavioral *** mechanism that makes NRXN1 a deletion hotspot is ***,we identified deletions of the NRXNlregion in case cohorts, confirming a strong association with autism spectrum and other neurodevelopmental disoMers Interestingly,deletions in both cases and controls were clustered in the 5'portionof NRXN1 and its immediate upstream *** explore the mechanism of deletion,we mapped and analyzed the breakpoints of 32 *** the deletion breakpoints,frequent microhomology(68.8%,2-19 bp) suggested predominant mechanisms of DNA replication error and/or microhomology-mediated end-joining and long terminal repeat(LTR) elements,unique non-B DNA structures and MEME-defined sequence motifs but not Alu and L1NE sequences were significantly enriched hnportantly,small-size inverted repeats(minus self chains,minus sequence motifs and partial complementary sequences) were significantly overrepresented in the vicinity of NRXNI region deletion breakpoints,suggesting that,while they are not interrupted by the deletion process,such inverted repeats can predispose to regional genomic instability by mediating single-strand DNA looping via the annealing of partially reverse complementary strands and promoting of DNA replication fork stalling and DNA replication *** observations highlight the potential importance of inverted repeats of variable size in generating a rearrangement hotspot in which individual breakpoints are not *** that involve short inverted repeats in initiating deletion may also apply to other deletion hotspots in the human genome.

关键词： NRXN1 exon-crossing deletion self chain sequence motif inverted repeat neurodevelopmental disorder

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Coronary heart disease is associated with a mutation in mitochondrial tRNA

Coronary heart disease is associated with a mutation in mito...

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者： JIA Zi-dong WANG Xin-jian QIN Yan-wen XUE Ling JIANG Ping-ping MENG Yan-zi SHI Su-xue WANG Yan MO Jun-qin Guan Min-xin Department of Genetics College of Life SciencesZhejiang University Division of Human Genetics Cincinnati Children's Hospital Medical Center Beijing Institute of Heart Lung and Blood Vessel Diseases Beijing Anzhen Hospital Affiliated to Capital Medical University Attardi Institute of Mitochondrial Biomedicine Wenzhou Medical University Department of Genetics College of Life SciencesZheJiang University Division of Pathology Cincinnati Children's Hospital Medical Center Division of Human Geneties Cincinnati Children's Hospital Medical Center

Coronary heart disease（CHD） is the leading cause of death *** genetic determinant for the development of CHD remains poorly *** report there the clinical,genetic,molecular and biochemical characterization of a four-generation Chinese family with maternally inherited *** of 32 adult members in this family exhibited variable severity and age-at-onset of *** analysis of their mitochondrial genomes identified the tRNA 15927G>A mutation belonging to the Eastern Asian haplogroup *** anticipated destabilization of a highly conserved base-pairing（28C-42G） by the 15927G>A mutation affects structure and function of *** analysis revealed≈80%decrease in the steady-state level of tRNA in the mutant cell lines carrying the 15927G>A *** 15927G>A mutation changed the conformation of tRNA,as suggested by slower electrophoretic mobility of mutated tRNA with respect to the wild-type *** addition,～39%reduction in aminoacylated efficiency oftRNA was observed in mutant cells derived from this Chinese family An in vivo mitochondrial protein labeling analysis showed～53%reduction in the rate of mitochondrial translation in mutant *** impaired mitochondrial protein synthesis leads to defects in overall respiratory capacity or malate/glutamate-promoted respiration or succinate/ glycero1-3-phosphatepromoted respiration,or N,N,N',N'-tetramethyl-pphenylenediamine/ ascorbate-promoted respiration in mutant *** increasing production of reactive oxygen species was observed in the mutant cells carrying the 15927G>A *** results provide the direct evidence that the tRNA 15927G>A mutation is associated with *** findings may provide new insights into pathophysiology and intervention targets of this disorder.

关键词： Coronary heart disease mitochondrial tRNAThr 15927G>A mutation

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Identification of substrate specificity of diacylglycerol acyltransferase 1(MucDGAT1)encoded by a cDNA isolated from developing cat's claw seed

Identification of substrate specificity of diacylglycerol ac...

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者： MAO-Xue WU Yong-Mei XUE Jin-Ai GAO Chang-Yong LI Run-Zhi Institute of Molecular Agriculture & Bioenergy Shanxi Agricultural University

Diacylglycerol acyltransferase(DGAT) catalyzes the acyl-CoA-dependent acylation of sn-1,2-diacylglycerol(DAG) to form seed oil triacylglycerol(TAG).There are two distinct types of non-homologous DGAT gene family designed as DGAT1 and DGAT2 encoding proteins with DGAT activity in plants and *** data evidence indicates that DGATs is one of the rate-limiting steps in plant lipid accumulation and plays an essential role in controlling both the quantitative and qualitative flux of fatty acids into storage *** DGAT substrate specificity,particularly for unusual fatty acids remains *** this work,a cDNA clone was isolated from developing seeds of cat's claw(Macfadyena unguis-cati),a naturally high accumulator of an unusual fatty acid palmitoleic acid(16:1△9).This cDNA was predicted to encode a DGAT1,named as ***1 expression was positively coordinated with palmitoleic acid and total oil accumulation in the developing *** expression system and in vitro enzyme activity assay combined with[14C]labeling substrates including DAG and Acyl-CoAs were employed to identify the substrate specificity of *** enzyme protein displayed a distinct substrate specificity for palmitoleate(Fig1).Furthermore,the co-expression of this enzyme and acyl-A9 desaturase(ScA9D) specifically in soybean seed led to palmitoleic acid increase by 2 folds more than palmitoleic acid level in the seeds of acyl-A9 desaturase expression alone(Fig 2).The present result was the first to demonstrate that MucDGAT1 can selectively incorporate palmitoleate into TAG,and functions crucially in high accumulation of 16:1△9 in plant ***,DGAT1 is one of important targets in metabolic engineering of common oil crops for commercial production of high-valued unusual fatty acids.

关键词： Identification of substrate specificity of diacylglycerol acyltransferase 1 MucDGAT1)encoded by a cDNA isolated from developing cat's claw seed

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Assembly of ω-7 fatty acid biosynthesis pathway in tobacco vegetable organs for high-quality biodiesel production

Assembly of ω-7 fatty acid biosynthesis pathway in tobacco ...

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者： GAO Chang-Yong GAO Xiu-Qing HUANG Chun-Guo ZHANG Li WANG Ji-Ping JIA Xiao-Yun YANG Zhi-Rong WU Yong-Mei MAO Xue LI Run-Zhi Institute of Molecular Agriculture & Bioenergy Shanxi Agricultural University

Biofuel oil is a stable,low carbon,renewable liquid fuel for direct fossil fuel replacement or blending *** use of common oil crops such as soybean and canola grown for biodiesel production results in more concerns on global food ***, development of alternative feedstock to replace food crops is crucial for wide-scale biofuel oil production to meet market *** is such a non-food biofuel crop when grown for energy production instead for smoking because tobacco can generate a huge amount of inexpensive biomass more efficiently than almost any other agricultural *** this work,we explored metabolic pathway assembly ofω-7 fatty acid biosynthesis in tobacco green tissues since oils rich in those fatty acids are the desirable sources for the production of high-quality biodiesel with much strong oxidative stability and cold tolerance,ω-7 fatty acids include palmitoleic acid(16:1 n-7) and its elongated products vaccenic acid(18:1 n-7) and eicosenoic acid(20:1 n-7).These fatty acids are not produced naturally in tobacco *** isolated and characterized the key enzyme genes responsible for palmitoleic acid synthesis from the high accumulators ofω-7 fatty acids such as macadamia(Macadamia integrifolia) and cat's claw(Dolichandra unguis-cati).Theω-7 fatty acid biosynthesis pathway was then engineered in tobacco by an integrative approach,which included co-overexpression of both cytosolic and plastid C16 specific n-7 fatty acid desaturase genes and RNA interference(RNAi) suppression of FATB C l6-ACP thioesterase *** to the wild-type tobacco where noω-7 fatty acids were produced,the transgenic tobacco plants synthesized and accumulatedω-7 fatty acids up to 40%in leaves and ***, palmitic(C16:0) and linoleic(C18:2) and a-linolenic(C18:3) acids were *** transgenic plants showed no significant changes in growth and development as well as other agronomic *** novel transgenic traits w

关键词： Assembly of fatty acid biosynthesis pathway in tobacco vegetable organs for high-quality biodiesel production

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DDF1调控水稻的营养生长与花器官发育

DDF1调控水稻的营养生长与花器官发育

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者：段远霖李生平陈志伟郑磊磊刁志娟周元昌兰涛官华忠潘润森薛勇彪吴为人作物遗传、育种与综合利用教育部重点实验室福建省水稻分子标记辅助育种重点实验室福建农林大学

Recent studies have shown that F-box proteins constitute a large family in eukaryotes and play pivotal roles in regulating various developmental processes in ***,their functions in monocots are still *** this study,we characterized a recessive mutant dwarf and deformedflowerl-1(ddf1-1) in *** mutant is abnormal in both vegetative and reproductive development,with significant size reduction in all organs except ***1 controls organ size by regulating both cell division and cell *** the ddf1-1 spikelet,the specification of floral organs in whorls 2 and 3 is altered,with most lodicules and stamens being transformed into glume-like organs and pistil-like organs,respectively;but the specification of lemma/palea and pistil in whorls 1 and 4 is not ***1 encodes an F-box protein anchored in nucleolus and is expressed in almost all vegetative and reproductive *** with the mutant floral phenotype,DDF1 positively regulates B-class genes OsMADS4/16,and negatively regulates pistil specification gene *** addition,DDF1 also negatively regulates the Arabidopsis LFY ortholog APO2,implicating a functional connection between DDF1 and ***,these results revealed that DDF1,as a newly identified F-box gene,is a crucial genetic factor with pleiotropic functions for both vegetative growth and floral organ specification in *** findings provide additional insights into the molecular mechanism controlling monocot vegetative and reproductive development.

关键词：水稻 ddf1 F-box 营养生长花器官发育

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RNA interference (RNAi) toward insect herbivores

RNA interference (RNAi) toward insect herbivores

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者： Hada Wuriyanghan Suvud Tong College of Life Sciences Inner Mongolia University Hohhot Inner Mongolia PR China.010021

RNA interference (RNAi) is a natural gene regulation and antiviral defense system of *** functions as a "gene silencing" mechanism by targeting specific RNA sequences for degradation (or possibly translational arrest) resulting in eliminating or reducing gene expression of the RNA *** are already several examples and even practical implementation of RNAi-based technologies for ***, RNAi technology has been proposed as a potentially useful tool for plant-associated insect control.

关键词： insect herbivore RNA interference transgenic plant

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