结构基因

In this article, the domain structure of LBD genes, classes of LBD genes in model plants of monocotyledon or dicot, expression model, mutation phenotypes from loss of function or gain of function, and its relationship to other gene families were reviewed. Furthermore, the redundancy character of LBD genes, basic candidate function shared by LOB domain, and potential molecular mechanism of LBD genes interacting with other genes also were discussed.

本文对LBD基因的结构域特征、在单／双子叶模式植物中的分类、表达特点、己克隆LBD基因的功能及与其它基因或基因家族间的相互作用关系进行了综述，并对LBD基因在高等植物中的功能冗余特性、LOB结构域可能所具有的基本功能和LBD基因与其它基因相互作用的分子机制进行了探讨。

Homology modeling of 3-D structure of two AChE from L.entomophila were constructed using H.sapiens(1p0i:A) native BuChE structure and Drosophila melanogaster(1d×4:A) native AChE structure as templates,respectively,by SWISS-MODEL.The catalytic triad were found and denoted in the 3-D structure of AChE from L. entomophila referring to T.californica.2.2 Gene cloning ofβ-actin and mRNA expression levels of two AChE genes from L. entomophilaBecause no reference gene has ever been used in Real Time PCR for L.entomophila in GeneBank,a fragment ofβ-actin gene was cloned from L.entomophila(GenBank Accession No.: FJ041117).It consists of 822 bp encoding a protein of 273 amino acids residues.

利用蛋白质结构同源建模工具，分别以人丁酰胆碱酯酶(1p0i：A)和果蝇乙酰胆碱酯酶(1d×4：A)的蛋白晶体结构为模板，对嗜虫书虱2个AChE的三维结构进行同源建模，并在三维结构中发现了AChE的酶解活性位点，证明嗜虫书虱体内也存在2个AChE基因。2.2嗜虫书虱β-actin基因克隆及乙酰胆碱酯酶基因mRNA表达水平研究目前关于嗜虫书虱的分子生物学研究较少，在GenBank中没有可用作内参基因的序列，因此本研究从嗜虫书虱体内克隆获得β—actin基因片段(GenBank登录号：FJ041117)，该片段长度为822 bp，编码273个氨基酸残基，同源性比对分析表明该片段与其它昆虫的β—actin基因具有很高的同源性。

In our study, 3 genotypies have been respectively demonstrated in UL139 and UL149 for the first time, and the corelations between certain genotypical structure and certain diseases were also proposed; The existence of the 3 genotypes of UL144 was foremost verified in isolates from congenital infants; Furthermore, the UL140, UL141 and UL145 genes were observed to be greatly discrepant to those had been described previously: comparing with Toledo, 231 nt are inserted in UL140, 2 more ORFs are obtained in UL141, and the UL145 ORF moves upstream by 90 nt. Except UL144, the sequences of 18 else genes in clinical isolates were submitted for the first time, and 479 sequences were assigned by GeneBank in total. Relevantly, 9 papers were published.

我们在学术界首次证实了HCMV UL139、UL149两个基因在临床分离株中分别存在三种基因型，发现了其中某个基因型的基因结构与特定来源的分离株存在一定的对应关系；首次在先天感染分离株中验证了UL144三种基因型的存在；首次证实了UL140、UL141和UL145等基因与原先认识的基因结构不同，其中UL140基因较Toledo株增加了231个碱基、UL141产生2个新的基因编码区、UL145 ORF较Toledo株前移90个碱基；首次或最先提交了除UL144基因外其余18个基因的临床分离株序列，本项目组共有479个HCMV相关基因序列被GenBank收录；发表研究论文9篇。

In this study, the genomic structure and regulatory elements of ApoD from 10 representive species including protozoan, invertebrate, protochordate and vertebrate were analyzed and compared. The genomic structure of ApoD is less conserved in organisms from protostome to deuterostome invertebrates, while it is highly conserved among chordates including amphioxus and verebrates. All four conserved cysteine residues are present in amino acid sequence of deuterostome ApoDs, while there are only two cysteine residues in amino acid sequence of protostomes ApoD. Structure divergence between protostome and deuterostome ApoD proteins suggests their function difference. The majority of regulatory elements are present in nearly all organism ApoD genes ranging from unicellular protozoan to mammals, suggesting that ApoD plays a very fundamental role, and possesses a conserved regulatory mechanism. However, there also exist some specific regulatory elements, which are present only in certain species and may perform some special roles.ApoD mRNA expression in murine NIH/3T3 fibroblasts exposed to various stresses such as as hydrogen peroxide and UV light shows dose-dependent increase. And a fly homolog of ApoD, Glial Lazarillo, whose overexpression results in increased resistance to hyperoxia as well as a extension of lifespan under normoxia and resistance to starvation without altering lipid or protein content.

本文首先从生物信息学角度对分属于原生动物、无脊椎动物、头索动物和脊椎动物类群的10种动物ApoD的基因结构及调控区的调控元件进行分析及比较，发现：(1)ApoD基因外显子-内含子结构从原生动物草履虫到原口动物再到后口动物海胆的进化过程中不保守，但在分析的几种脊椎动物中相当保守；(2)文昌鱼ApoD基因扮演从无脊椎动物到脊椎动物承上启下的角色，可能代表了脊椎动物ApoD基因原型；(3)四个半胱氨酸保守位点在后口动物中都存在，而在原口动物中只存在两个，原口、后口动物ApoD蛋白一级结构上的差异反映蛋白功能上可能也存在一定差别；(4)调控区大多数主要调控元件为不同动物共有，说明ApoD主要功能及其表达调控在进化中相当保守；(5)ApoD基因个别调控元件是随着物种进化而出现并开始发挥相关作用，如SF-1；还有一些调控元件在进化过程中还没有发现其规律，这说明ApoD某些功能和基因表达调控模式可能因物种不同而存在一定的差异。

Attenuation The trp operon is also controlled by attenuation A leader sequence in the polycistronic mRNA upsteam of the coding region of the trpE structural gene encodes a 14 amino acid leader peptide including two tryptophan residues The RNA leader sequence can for several possible stem-loop secondary structures,one of which can act as a transcription terminator whilst a different stem-loop can act as an anti-terminator.

弱化作用 trp 操纵子也受到弱化作用的调控。在trpE结构基因编码区上游的多顺反子mRAN上有一段前导序列，可以编码一个含有14个氨基酸的前导肽，此肽中含有两个色氨酸残基。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫，第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株；随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛，三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接／酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D、pTARGET／P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1／IFN分别／同时免疫，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：随后将质粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1／IFN同时免疫牛，三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

Objective:To investigate the relationship between drug resistance and the mutant of ampC structure gene in Enterobacter cloacae.

目的 ：探讨阴沟肠杆菌的耐药性与ampC结构基因突变之间的关系。

C The structural genes are not contained in operons.

操纵组中不含结构基因

Random sampling 36 wild Japanese quails from which migrated to and settled down the Weishan Lake area,and the gene frequency of 10 loci in viscera and muscle is detected. After collecting the same data about the 20 quail colonies in China and other populations and applying fuzzy discriminants, the domestic quails is discriminated to belong to which populations, wild Japanese quails or wild quails in the Weishan Lake area.

在迁飞滞留季节捕获的微山湖地区野生鹌鹑中随机抽取36只，检测编码脏器、肌肉酶型的10个结构基因座上的频率分布，搜集国内外20个家鹑和野生日本鹌鹑群体的相同资料，应用模式判别方法，以家鹑群体为识别对象，分别与野生日本鹌鹑和微山湖野生鹌鹑群体进行模糊性识别。

Chrysanthemum of 10 thousand birthday is lax to edaphic requirement, with the arenaceous qualitative loam with fecund, good drainage had better.

万寿菊对土壤要求不严，以肥沃。排水良好的砂质壤土为好。

He unstepped the mast and furled the sail and tied it.

他拔下桅杆，把帆卷起，系住。