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K562细胞未知基因KH的开放阅读框架在大肠杆菌中的表达

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重庆医科大学学报 2004年第2期29卷 124-126页

作者：何於娟欧一衡张彦刘小珊马凌娣涂植光蒋纪恺重庆医科大学基础医学院临床生化教研室重庆400016 重庆市永川市人民医院重庆402160 重庆医科大学临床学院血液科重庆400016

目的 :构建K5 6 2细胞未知基因KH的开放阅读框架 (openreadingflame ,ORF)的原核表达体系。方法 :采用DNA重组技术构建KH基因片段的开放阅读框架 (KH -ORF)的表达载体pCI-neo-KH -ORF ,热休克法转化大肠杆菌BL2 1(DE3)plysS ,IPTG诱导... 详细信息

目的 :构建K5 6 2细胞未知基因KH的开放阅读框架 (openreadingflame ,ORF)的原核表达体系。方法 :采用DNA重组技术构建KH基因片段的开放阅读框架 (KH -ORF)的表达载体pCI-neo-KH -ORF ,热休克法转化大肠杆菌BL2 1(DE3)plysS ,IPTG诱导后 ,SDS -PAGE鉴定目的蛋白的表达。结果 :经IPTG诱导 0 .5h后KH -ORF在BL2 1(DE3)plysS中获得表达 ,表达产物以包涵体形式存在 ,分子产量约 2 4kD。结论 :成功构建了KH -ORF的原核表达体系 ,为进一步研究其功能奠定了基础。

关键词： K562细胞 KH基因开放阅读框架原核表达

一含有两个开放阅读框架的cDNA片段的发现(英文)

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免疫学杂志 2000年第5期16卷 321-324页

作者：姜颖于永利白求恩医科大学免疫学教研室吉林长春130021

目的分离人知的 c DNAs。方法采用 RT- PCR自人的肝脏细胞分离到一约 1,2 0 0 bp的 c DNA片段 ,将其克隆测序。结果在这一 c DNA片段中包含了两个开放阅读框架。结论一个开放阅读框架代表与人疱疹病毒大衣壳蛋白同源的多肽 ,另一个代表... 详细信息

目的分离人知的 c DNAs。方法采用 RT- PCR自人的肝脏细胞分离到一约 1,2 0 0 bp的 c DNA片段 ,将其克隆测序。结果在这一 c DNA片段中包含了两个开放阅读框架。结论一个开放阅读框架代表与人疱疹病毒大衣壳蛋白同源的多肽 ,另一个代表与沙门氏菌外膜蛋白

关键词： DNA片段血管生成抑制开放阅读框架多肽

蛋白质组学发现结核分枝杆菌H37Rv基因组学未预测的开放阅读框架

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国外医学（微生物学分册） 2002年第6期25卷 43-44页

作者：乐军

每年结核病的新发病例为800万,病死人数200万,世界卫生组织宣布全球结核病处于紧急状态,亟需预防和治疗结核病的新策略。蛋白质组学反映细胞对环境刺激的功能状态,因此它可作为基因组学有价值的补充。为了寻找免疫干预的新策略,通过比... 详细信息

每年结核病的新发病例为800万,病死人数200万,世界卫生组织宣布全球结核病处于紧急状态,亟需预防和治疗结核病的新策略。蛋白质组学反映细胞对环境刺激的功能状态,因此它可作为基因组学有价值的补充。为了寻找免疫干预的新策略,通过比较结核分枝杆菌(MTB)有毒株与无毒疫苗株之间蛋白质组成的差异进行系统蛋白质组的研究。

关键词：蛋白质组学结核分枝杆菌 H37Rv 开放阅读框架

Pod1 is involved in the sexual differentiation and gonadal development of the Nile tilapia

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Science China(Life Sciences) 2011年第11期54卷 1005-1010页

作者： LI JianZhong LIU ShaoJun NAGAHAMA Yoshitaka Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education College of Life Sciences Hunan Normal Univer sity Changsha 410081 China Laboratory of Reproductive Biology National Institute for Basic Biology Okazaki 444-8585 Japan

Pod1 is a member of the basic helix-loop-helix(bHLH) family of transcription factors that have been implicated in the regulation of sexual differentiation and gonadal development in mammals.However,to date,little is known about the role of Pod1 in nonmammalian vertebrate gonadogenesis.We cloned and characterized the Pod1 gene from tilapia.The tilapia Pod1 gene contains an open reading frame(ORF) of 525 nucleotides which potentially codes for a protein with 174 amino acids.Sequence alignment revealed that the deduced tilapia protein sequence shared high homology(79.5% to 90.5%) with the Pod1 sequences of other vertebrates.The tissue distribution of Pod1 revealed by RT-PCR showed that it had varied expression patterns in adult tilapia.In situ hybridization was performed to examine the temporal and spatial expression patterns of Pod1 during tilapia sexual differentiation and gonadal development.In the undifferentiated gonad,Pod1 was expressed in the somatic cells of both sexes.Subsequently,Pod1 expression in tilapia persisted in differentiated juvenile and adult ovary and testis.Our data indicate for the first time that Pod1 is not only necessary for the onset of sexual differentiation,but also plays an important role in gonadal development in the teleost.

关键词：尼罗罗非鱼性腺发育性别分化 RT-PCR分析蛋白质序列开放阅读框架哺乳动物脊椎动物

Polymorphisms of Epstein-Barr virus BHRF1 gene, a homologue of bcl-2

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Chinese Journal of Cancer 2010年第12期29卷 1000-1005页

作者： Yong-Zheng Jing Yun Wang Yu-Ping Jia Bing Luo Department of Microbiology Qingdao University Medical College Qingdao Shandong 266021 P. R. China Department of Clinical Laboratory Municipal Hospital of Penglai Penglai Shandong 265600 P. R. China

Background and Objective: EBV BamHI fragment H rightward open reading frame 1 (BHRF1), the Epstein-Barr virus (EBV) early gene, is structurally and functionally homologous to the oncogene bcl-2 and may play an important role in the development of EBV-associated tumors. To characterize the polymorphisms of BHRF1 in EBV-associated tumors, we analyzed the sequences of BHRF1 in isolates from nasopharyngeal carcinoma (NPC) and EBV-associated gastric carcinoma (EBVaGC) biopsies as well as throat washing (TW) samples from healthy donors. Methods: BHRF1 DNA sequences were analyzed by polymerase chain reaction (PCR) and sequencing for 39 NPC samples, 40 EBVaGC samples, and 53 EBV-positive TW samples from healthy donors. The variants of BHRF1 gene were classified according to the signature changes. The EBV types 1 and 2 at nuclear antigen (EBNA) 3C locus were determined by PCR. Results: Compared with EBV standard cell line B95-8, all isolates carried a silent mutation at amino acid (AA) 80 (nucleotide 54616 T→C), the AA88 L→V mutation was found in most isolates, and the AA79 V→L mutation in a few isolates. Other mutations were sporadically distributed. Based on the mutations at AA88 and AA79, 3 distinct variants of BHRF1 genes, designated as 79V88V, 79L88L, and 79V88L, were identified. The 79V88V was the most common variant. The distribution of the BHRF1 variants among the NPC, EBVaGC, and TW samples was not significant. The corresponding regions of bcl-2 homologues were conserved in all isolates except for 3 samples. The distribution of BHRF1 variants in type 1 and type 2 strains was significant different (P < 0.001, contingency coefficient was 0.554). Conclusions: The 79V88V is the dominant variant in NPC, EBVaGC, and TW samples from healthy donors and preferential linkages between BHRF1 and EBNA3C variants exist. Conserved BHRF1 in Bcl-2 homologous domains is helpful to remain the important role of BHRF1.

关键词： EB病毒早期基因同源基因多态性全国人民代表大会基因突变开放阅读框架聚合酶链反应

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真核生物中的“双重编码”现象

生物化学与生物物理进展 2009年第5期36卷 536-540页

作者：申志勇李稚锋杭兴宜张成岗军事医学科学院放射与辐射医学研究所蛋白质组学国家重点实验室北京100850

双重编码(dual coding)是指一段mRNA序列同方向上存在两个开放阅读框架,从而可编码不同氨基酸序列的现象.双重编码现象在原核生物、噬菌体和病毒中作为一种可有效利用基因组的方式而普遍存在.新近报道,在真核生物中也存在双重编码现象,... 详细信息

双重编码(dual coding)是指一段mRNA序列同方向上存在两个开放阅读框架,从而可编码不同氨基酸序列的现象.双重编码现象在原核生物、噬菌体和病毒中作为一种可有效利用基因组的方式而普遍存在.新近报道,在真核生物中也存在双重编码现象,对于认识真核生物基因表达调控的机制提供了新的信息.

关键词：双重编码基因开放阅读框架真核生物

Human Cytomegalovirus UL138 Open Reading Frame Is Highly Conserved in Clinical Strains

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Chinese Medical Sciences Journal 2009年第2期24卷 107-111页

作者： Ying Qi Rong He Yan-ping Ma Zheng-rong Sun Yao-hua Ji Qiang Ruan Virus Laboratory Shengjing Hospital China Medical University Shenyang 110004 China

Objective To investigate the variability of human cytomegalovirus(HCMV) UL138 open reading frame(ORF) in clinical strains.Methods HCMV UL138 ORF was amplified by polymerase chain reaction(PCR) and PCR amplification products were sequenced directly,and the data were analyzed in 19 clinical strains.Results UL138 ORF in all 30 clinical strains was amplified successfully.Compared with that of Toledo strain,the nucleotide and amino acid sequence identities of UL138 ORF in all strains were 97.41% to 99.41% and 98.24% to 99.42%,respectively.All of the nucleotide mutations were substitutions.The spatial structure and post-translational modification sites of UL138 encoded proteins were conserved.The result of phylogenetic tree showed that HCMV UL138 sequence variations were not definitely related with different clinical symptoms.Conclusion HCMV UL138 ORF in clinical strains is high conservation,which might be helpful for UL138 encoded protein to play a role in latent infection of HCMV.

关键词：人巨细胞病毒开放阅读框架临床症状菌株 PCR扩增产物编码蛋白质保守聚合酶链反应

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猪ILF2基因的电子克隆与生物信息学分析

江苏农业科学 2011年第4期39卷 23-26页

作者：孙婴宁王维禹齐齐哈尔大学生命科学与农林学院黑龙江齐齐哈尔161006

白介素增强子结合因子2调节白介素2基因表达的转录过程,属于重要的免疫相关基因。利用电子克隆技术,以人的ILF2基因(NM_004515)为种子序列,克隆了猪的ILF2基因。结果显示该cDNA序列长1 628 bp,包含1条长度为1 173 kb的开放阅读框架,5′... 详细信息

白介素增强子结合因子2调节白介素2基因表达的转录过程,属于重要的免疫相关基因。利用电子克隆技术,以人的ILF2基因(NM_004515)为种子序列,克隆了猪的ILF2基因。结果显示该cDNA序列长1 628 bp,包含1条长度为1 173 kb的开放阅读框架,5′非编码区长90 bp,3′非编码区长365 bp,编码390个氨基酸。推导的氨基酸序列与牛、马、人、小鼠、非洲爪蟾、斑马鱼的同源性分别为99%、99%、99%、99%、90%、87%。理论等电点为5.19,分子量为43 050.12 u。N端含有一个富含RGG的单链RNA结合基序,C端含有1个DZF锌指结构。猪ILF2基因的成功克隆,为进一步研究其基因结构、基因表达与调控奠定了基础。

关键词：白介素增强子结合因子2 白介素2 电子克隆生物信息学表达序列标签开放阅读框架

Construction and Identification of the Helper Plasmids for Reverse Genetic System of Rabies Virus Street Strain

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Virologica Sinica 2009年第6期24卷 559-565页

作者： Ping-gang MING Ying HUANG Qing TANG Jia-liang DU Xiao-yan TAO Jia-xin YAN Rong-liang HU State Key Laboratory for Molecular Virology and Genetic Engineering Institute for Viral Disease Control and Prevention Chinese Center for Disease Control and Prevention （IVDC China CDC） Beijing 100052 China Department of Genetic Engineering Wuhan Institute of Biological Products Wuhan 430060 China Laboratory of Epidemiology Veterinary Institute Academy of Military Medical Sciences Changchun 130062 China

To obtain the helper plasmids for a reverse genetics system of rabies virus, the cDNAs of the complete open reading frames of the N, P, G, and L genes of rabies street virus stain HN10 were each cloned into expression vector pVAX1, These four plasmids were identified by restriction enzyme digestion and gene sequencing. The plasmid encoding the N protein was selected to determine the expression effect of these plasmids in NA cells. The results showed that the helper plasmids for a reverse genetics system of rabies street virus strain HN10 had been successfully constructed.

关键词：狂犬病病毒反向遗传学质粒表达应变系统鉴定开放阅读框架基因测序狂犬病毒