基因的

In this experiment, we screen the major protective antigen gene-SOD gene of M. paratuberculosisin order to study the sensitive, specific diagnostic reagent and prophylaxis preparation, especially theDNA vaccine. The SOD gene was amplified from Mycobacterium paratuberculosis C-2 chromosomalDNA by using the PCR technique and cloned into pMD18-T Vector System. We gained a SOD gene of624bp.The recombinant clone was identified byα-complementarity, enzyme digestion and PCRidentification. The result indicated that the recombinant plasmid pMD18-T-SOD was successfullyconstructed. Moreover, through sequential determination and DNASTAR analysis between the clonedSOD gene of M. paratuberculosis C-2 and that of the M.paratuberculosis K-10 strain, the sequentialhomogeneity reached 99%, and the amino acid homogeneity reached 99.5%. The preceding analysisindicated that the SOD gene was very conservative in M. paratuberculosis.

为了研制副结核病敏感、特异的诊断试剂和新型、高效的预防制剂，尤其是DNA疫苗，本研究筛选了M.paratuberculosis主要保护性抗原SOD基因，以M.paratuberculosis C-2染色体DNA为模板，以SOD基因的特异性引物进行PCR扩增，获得了624bp的SOD基因，通过T-A克隆技术，将PCR产物克隆至pMD18-T Vector中，以质粒大小、酶切分析、PCR扩增及序列分析鉴定重组克隆，成功地构建出克隆质粒pMD18-T-SOD，序列测定及DNASTAR分析表明，所获得的M.paratuberculosis C-2 SOD基因与Gen Bank中M.paratuberculosis K-10 SOD基因的大小完全一致，两者核苷酸序列的同源性为99％，氨基酸序列的同源性为99.5％，表明该基因在副结核分枝杆菌中是高度保守的。

It also can be used as alternative antigen of newgeneration vaccine.In this experiment,we screen the major protective antigen hsp65 gene of MAP in order to developnew vaccine especially the DNA vaccine for the prevention of paratuberculosis disease.The hsp65 genewas amplified from MAP C-2 chromosomal DNA by using the PCR technique.We gained a hsp65 gene of 1 626bp.Then PCR product was cloned into pGEM-T vector by T-A clone technique and therecombinant clone was identified by plasmid size,enzyme digestion and PCR identification.The cloneplasmid of pGEM-T- hsp65 was successfully constructed.The nucleotide sequence and deduced aminoacid sequence ofclone gene was analyzed by DNASTAR software.The result indicated that the size ofhsp65 gene consist with M.paratuberculosis K-10 strain in GenBank and the sequential homogeneityreached 99.1%,the amino acid homogeneity reached 99.3%.The preceding analysis indicated that thehsp65 gene was very conservative in M.paratuberculosis.

为了研发预防副结核病的新型疫苗尤其是DNA疫苗及相关蛋白功能，本研究选择了MAP的主要保护性抗原Hsp65蛋白，以副结核分枝杆菌C-2株染色体DNA为模板，以hsp65基因的特异性引物进行PCR扩增，获得了1 626bp的hsp65基因，通过T-A克隆技术，将PCR产物克隆至pGEM-T Vector中，以质粒大小、酶切分析、PCR扩增及序列分析鉴定重组克隆，成功地构建出克隆质粒pGEM-T-hsp65，以DNASTAR软件分析了所克隆基因的核苷酸序列和推导的氨基酸序列，结果表明，所获得的hsp65基因与GenBank中MAPK-10株该基因核苷酸大小完全一致，两者核苷酸序列的同源性为99.1％，氨基酸序列的同源性为99.3％，表明该基因在副结核分枝杆菌中高度保守。

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。

Sequences of modified genes GOX and CP4-EPSPS in GM canola and that of modified gene Cryla in GM cotton were decoded and the conservative sequences of exogenous resistant genes: PLRVrep, PVYcp and CryⅢA in GM potato: New leaf〓 PLUS and New leaf〓 Y were confirmed using the modified exogenous gene decoding technique and isogenous sequence similarity BLAST analysis. Bases on DNA sequences studied above, conventional PCR primers were designed and selected in an optimized way and the conventional PCR detection protocol for exogenous resistant genes in 19 GM crops was established.

本研究采用反向PCR克隆—测序测定未知序列技术，针对转基因玉米（MON810、BT11、BT176、GA21、T25、CBH-351）六个品系、转基因大豆GTS 40-3-2品系、转基因油菜RT73和MS8两个品系、转基因棉花MON531和MON1445两个品系测定出品系鉴定的边界序列；采用外源修饰基因序列的破译方法和BLAST同源性序列分析技术，针对转基因作物中修饰的外源抗性基因进行破译研究，破译出转基因油菜中修饰的GOX基因、修饰的CP4-EPSPS基因和转基因棉花中修饰的Cry1A基因的序列，并确定了转基因马铃署New leaf〓PLUS和New leaf〓Y两个品系中PLRVrep、PVYcp和CryⅢA外源抗性基因保守序列。

Of the 32 articles, 4 were about the selection of destination genes, 3 about the selection of target cells, 7 about the selection of gene expression vector, 3 about the methods of gene transfer, 4 about gene therapy combined with tissue engineering in treating bone defect, 5 about polygenes in treating bone defects, and 2 about the problems and prospects in the gene therapy of bone defect.

符合纳入标准的32篇文献中，4篇涉及目的基因的选择，3篇涉及靶细胞的选择，7篇涉及基因表达载体的选择，3篇涉及基因转移方法，4篇涉及基因治疗与组织工程结合治疗骨缺损，5篇涉及多基因联合治疗骨缺损，2篇涉及骨缺损基因治疗存在的问题与展望。

Selecting the better amplified from Diacylglycerol acyltransferase, gene, Acyl- desaturase gene ,then cloning sequencing, afer that, through the NCBI and nucleotide sequence to match homology , the results show that: cloning by the cassava, castor-oil plant Jatropha curcas DGAT and SAD genes and gene fragments have the high homology with other known plant DGAT genes and SAD genes .

从中筛选出扩增效果较好的二酰基甘油酰基转移酶(Diacylglycerol acyltransferase， DGAT)基因，硬脂酰-酰基载体蛋白脱饱和酶(Acyl- desaturase， SAD)基因克隆测序，经测序，通过NCBI与已知的核苷酸序列进行同源性比对，结果表明：克隆所获得的木薯、蓖麻和麻疯树 DGAT基因和SAD基因的片段与其它已知植物的DGAT基因和SAD基因具有很高的同源性。

Selecting the better amplified from Diacylglycerol acyltransferase, gene, Acyl- desaturase gene ,then cloning sequencing, afer that, through the NCBI and nucleotide sequence to match homology , the results show that: cloning by the cassava, castor-oil plant Jatropha curcas DGAT and SAD genes and gene fragments have the high homology with other known plant DGAT genes and SAD genes .

从中筛选出扩增效果较好的二基甘油基转移(Diacylglycerol acyltransferase， DGAT)基因，硬脂-基载体蛋白脱饱和(Acyl- desaturase， SAD)基因克隆测序，经测序，通过NCBI与已知的核酸序列进行同源性比对，结果表明：克隆所获得的木薯、蓖麻和麻疯树 DGAT基因和SAD基因的片段与其它已知植物的DGAT基因和SAD基因具有很高的同源性。

Construct PLXRN-IL-1Ra and PLXRN-IL-10 vector, lapine synoviocytes were first transduced in culture by retroviral infection. The genetically modified synovial cells were then transplanted by intra-articular injection into the knee joints, assay of joint lavages confirmed that the gene expression was not lost after 14 days of transfer. Knees receiving the hIL-1Ra had significantly reduced cartilage breakdown. Delivery of the hIL-10 was less effective, When both genes were used together, there was a greater inhibition of cartilage breakdown and a considerable reduction of cartilage matrix degradation.

构建PLXRN-IL-1Ra、PLXRN-IL-10逆转录病毒载体，体外感染同种异体原代滑膜细胞，接着将转染了外源基因的滑膜细胞行关节腔注射入创伤性骨关节炎兔膝关节，通过对关节滑液的ELISA分析证明外源基因的表达在基因转移后14天还很稳定，只接受hIL-1Ra基因治疗的膝关节软骨损坏明显减轻，接受hIL-10基因治疗的膝关节治疗也有一定效果，当两种基因同时导入时，有非常明显的抑制软骨破坏和软骨基质降解的作用。

Construction of the homologous arms directing the homologous recombination For the sake of seeking for the more efficient integrating strategy to make the foreign gene insert in the specific site and don't destroy the host expressional and regulatory elements such as promoters and introns, we experimented targeting the Bombyx morl fibroin heavy-chain exon II with the green fluorescent protein gene by homologous recombination technology.

为了探索一条更为有效的整合策略，使外源基因能定点整合到靶位点，又不破坏宿主基因的启动子、内含子等表达调控元件，令外源基因对宿主生理功能的影响达到最小，我们以丝素重链基因2号外显子为整合靶位点，以便将外源基因定点整合其间。

Indica variety ZYQ8 is an important genetic resource resistant to rice blast disease in rice breeding in North China.in this study,resistance identification of the populations of F1,F2,DH and B1F1of the crosses of ZYQ8×Jingxi17 (JX17) and ZYQ8×Lijiangxintuanheigu was carried out by using a Chinese differential strainZH10-8-14 from North China and a Japanese differential strain Ken54-04 from Japan and the results demonstrated that ZYQ8 had one dominant major gene Pi-zh which had been reported by Zhu L.H., et al., The results from experiments in which the F2 populations of ZYQ8×nine different varieties with known resistant genes were inoculated with strain Ken54-04 indicated that the gene Pi-zh was a new gene and non-allelic to nine known gene loci,namely Pi-i,Pi-km,Pi-z,Pi-ta,Pi-ta2,Pi-zt,Pi-kp,Pi-b and Pi-t.

籼稻品种窄叶青8号是我国北方稻区水稻育种上重要的稻瘟病抗源之一。本文利用我国北方稻区的代表菌系中10-8-14和日本的代表菌系研54-04，对窄叶青8号与感病品种京系17号和丽江新团黑谷的杂交F1、F2、DH和B1F1群体进行抗病性鉴定。根据抗病性的分离，确认窄叶青8号的抗性由1对显性主效基因，即朱立煌等报道的Pi-zh基因控制。利用菌系研54-04接种窄叶青8号与9个具有已知抗病基因的鉴别品种杂交的F2群体，各群体都表现二基因的独立遗传，证明Pi-zh基因与Pi-i、Pi-km|、Pi-z、Pi-ta、Pi-taz、Pi-zt、Pi-kp、Pi-b和Pi-t等9个已知抗病基因间存在非等位关系，是新的抗稻瘟病基因。

However, as the name（read－only memory）implies, CD disks cannot be written onorchanged in any way.

然而，正如其名字所指出的那样，CD盘不能写，也不能用任何方式改变其内容。

Galvanizes steel pallet is mainly export which suits standard packing of European Union, the North America. galvanizes steel pallet is suitable to heavy rack. Pallet surface can design plate type, corrugated and the gap form, satisfies the different requirements.

镀锌钢托盘多用于出口，替代木托盘，免薰蒸，符合欧盟、北美各国对出口货物包装材料的法令要求；喷涂钢托盘适用于重载上货架之用，托盘表面根据需要制作成平板状、波纹状及间隔形式，满足不同的使用要求。