基因型

We can make use of chip to analysis virus gene group; construct mutation detection technique of heredity diseases and become normal detection technique of many heredity diseases; supervise the changes of cell gene expression, research pathogenic theory of virus; differentiate bacteria gene types and identify bacteria strains, provide base data for screening and supervise drug-fast gene.

利用基因芯片可加速对病毒基因组的功能分析；可建立遗传病的突变检测技术，并成为对多种遗传病的常规诊断技术；可用来监测宿主细胞基因表达的改变，研究病毒的致病机理；基因芯片还可用来对细菌基因的分型和菌种鉴定，为筛选和监测细菌的抗药性基因提供基础数据。

Strains containing cry1 genes were the most abundant in our collection (66%), and 21 different cry1-type gene combinations were found. Furthermore, several novel holotype cry genes were found and the full-length sequences of 3 novel cry genes were designated as cry54Aa1, cry30Fa1, and cry30Ga1 by B.

PCR-RFLP鉴定结果表明：此地区的苏云金芽胞杆菌主要含有cry1，cry2，cry3，cry4/10，cry9，cry30和 cry40 等7种cry基因类型；含cry1基因的菌株最丰富，共有21种不同cry1型基因组合；从中发现了新型模式基因，并采用Tail-PCR技术获得了其中3个基因的全长序列，被国际苏云金芽胞杆菌杀虫晶体蛋白基因命名委员会命名为cry54Aa1、cry30Fa1和cry30Ga1。

Coil O85 antigen contained 8 varieties of genes, including UDP-Nacetylglucosamine-2-epimerase gene (f1). UDP-galactopyranose mutase gene, glycosyl transferase genes (f3, f5, f6, f8), O-antigen transferase gene and O-antigen polymerase gene, in which two genes, wzx and wzy and 4 pairs of primers were identified to be specific to E.

发现8个开放阅读框架并确定功能，分别为：UDP-N-乙酰葡萄糖－2-异构酶基因，吡喃型UDP-半乳糖变位酶基因，糖基转移酶基因（orf3、orf5、orf6和orf8），O-抗原转运酶基因和O-抗原聚合酶基因。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体；2）利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达；3）构建ZNRD1的小干扰RNA载体，并测序鉴定；4）利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞（AGS、SGC7901、MKN28）和小鼠成纤维细胞（NIH3T3），G418筛选后进行鉴定；5）利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞（SGC7901）、正常胃组织上皮细胞（GES-1）、对长春新碱耐药的胃癌细胞（SGC7901/VCR）、药敏白血病细胞（HL-60）、对长春新碱耐药的白血病细胞（HL-60/VCR），G418筛选后进行鉴定；6）利用MTT实验检测ZNRD1高/低表达对细胞（AGS、SGC7901、MKN28、NIH3T3、GES-1）生长的影响；7）通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响；8）通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响；9）通过流式细胞仪分析ZNRD1高/低表达对细胞（AGS、MKN28、NIH3T3、GES-1）的细胞周期的影响；10）通过流式细胞仪分析上调ZNRD1对细胞（AGS、MKN28、NIH3T3）的凋亡的影响；11）通过MTT实验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）体外药物敏感性的影响；12）通过肾包膜下移植法检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR）体内药物敏感性的影响；13）通过流式细胞仪分析ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）内阿霉素蓄积和泵出的影响；14）通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响；15）通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60）凋亡敏感性的影响；16）通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响；17）利用RT-PCR、Western blot对基因芯片的结果进行鉴定；18）利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用；19）利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用；20）利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响；21）利用反义核酸技术抑制p21的表达；通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响；22）利用反义核酸技术抑制p27的表达；通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响；23）利用脂质体转染法上调Skp2的表达；通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响；24）利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响；25）利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响；26）利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响；27）利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞（SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR）多药耐药表型的影响；利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

The new GV preparation method we developed allowed the best visualization of chromatin distribution after Hoechst or CMA3 staining. Our results showed that GV chromatin configurations are different among different species, and thus, while chromatin condensed into a ring around the nucleolus in porcine and bovine oocytes, no such a ring was observed in caprine oocytes; Porcine oocytes were synchronized at GV1, goat oocytes at GV3n and bovine oocytes at GVf configurations before GVBD occurred, indicating that these configurations are progressive towards final maturation rather than atresia; Changes of chromatin configurations were associated with gene activity of transcription; GV chromatin configurations were affected by the environmental changes such as serum starvation and elevated temperature.

应用该技术对猪、山羊和牛不同大小、不同健康程度卵泡及体内和体外成熟过程中卵母细胞染色质构型的变化发现：1）不同动物卵母细胞染色质构型不同；猪和牛卵母细胞染色质围绕核仁凝集形成环，但山羊卵则没有此环。2）卵母细胞在GVBD之前都同步在某一染色质构型；猪同步在GV1，山羊GV3n，牛GVf，说明这些染色质构型代表着进行性变化。3）染色质构型的变化反映基因转录活动的变化。4）染色质构型受环境变化的影响。

In addition, baculovirus are noninfectious to vertebrates, and their promoters have been shown to be inactive in mammalian cells. Futhermore, the expression level of some recombinant proteins is up to 1000 mg/L. Finally, recombinant proteins are soluble and antigenically, immunogenically and functionally similar to their authentic counterparts. In this article, we used the transfer vector pBlueBac of AcNPV (Autographa californica Nuclear Polyhedrosis Virus) to construct the recombinant baculovirus AcNPV-GM-CSF by in vivo recombinantion.

本文中，我们首先利用苜蓿夜蛾核型多角体病毒带β-Galactosidase 基因标记的非融合蛋白基因转移载体pBlueBac，通过细胞内同源重组的方式将hGM-CSF基因成功地插入病毒AcNPV的基因组中。hGM-CSF基因编码信号肽和完整的氨基酸序列。hGM-CSF基因在感染重组病毒的草地夜蛾培养细胞Sf9中得到表达，感染后的Sf9细胞培养液能刺激人骨髓细胞在体外形成典型的集落，表达水平可达2.7〓CFU／ml。

To obtain more information of magnesium homeostasis in mitochondria, mTn-lacZ/LEU2 transposon library was transformed into mrs2 deletion mutant to screen for suppressor genes of MRS2. YMR166C, a member of mitochondrial carrier family, was identified as a suppressor gene of MRS2. Deletion of YMR166C gene can rescue the defects of mrs2 deletion mutant such as the decrease in mitochondrial magnesium concentration, Group II RNA splicing defect and growth defect on nonfermentable carbon source. For the first time we demonstrated YMR166C is involved in mitochondrial magnesium homeostasis.

为了增进对线粒体镁离子代谢调控基因的了解，利用酿酒酵母mTn-lacZ/LEU2转座子文库筛选MRS2的抑制基因，发现线粒体载体家族成员YMR166C基因的缺失可以挽救MRS2基因缺失的突变体的生长缺陷、II型内含子剪接缺陷，并可以调节线粒体镁离子浓度，首次发现YMR166C是线粒体镁代谢相关基因。

Personality traits were evaluated by Eysency Personality Questionnaire that was compiled by Eysency and revised by Gong. Standard scores of Psychoticism, Extraversion and Neuroticism were calculated, and taking 50 points as a boundary. Each dimension was assigned into 2 grades non-psychotic type (P0) psychotic type (P≥50), introversion type (E0), extraversion type (E0), non-nervous type (N0) and nervous type (N≥50). 5 mL ulnar vein blood was obtained without adding decoagulant, and then serum was removed. Genome DNA was extracted from blood clot by conventional chloroform for PCR analysis. PCR product was sequenced by pyrophosphoric acid. SNP was determined by PSQ96 MA sequenator. Allele frequency was analyzed by mass spectrometry.

人格评定采用Eysency编制，龚耀先修订的艾森克人格问卷、计算精神质，内外问，神经质3个个性维度的标准分，并以50分为界，将各维度分为两个等级：非精神病倾向型（精神质0）、精神病倾向型（精神质≥50）；内倾型（内外向0），外倾型（内外向≥多50）；非常经质倾向型（神经质0），神经质倾向型（神经质≥50）取肘静脉血5 mL，不加抗凝剂，弃血清，血凝块常规酚氯仿提取基因组DNA，进行PCR分析PCR产物应用焦磷酸测序PSQ96 MA测序仪进行SNP检测，用质谱法进行及等位方基因频率分析。

To obtain the tervalent fusion toxin gene, three toxin gene fragments from three species of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio mimicus were connected with the flexible linker using overlap extension PCR. The three toxin gene fragments respectively encode the mature proteins of the thermostable direct hemolysin of V. parahaemolyticus, the cytotoxin of V. vulnificus and the heat-labile hemolysin of V.

采用柔性Linker和重叠延伸PCR方法将三个毒素基因，即副溶血性弧菌的耐热型直接溶血素基因tdh、创伤弧菌的溶细胞毒素基因vvhA和拟态弧菌的热不稳定型溶血素基因vmhA的成熟蛋白基因片段进行串联，得到三联融合毒素基因tdh-vvhA-vmhA。

The RhoA oncogene , have been revealed in recent years, are closely related to various malignant tumours. RhoA, belonging to Rho family, is one member of the ras superfamily, whose encoding production, namely RhoA protein, pertaining to micromolecular mass GTP conjugated protein family, in GTP conjunctive active form and GDP conjunctive unreactive form. It exist in the latter form in normal cells and the switch of the two forms enables them to educe functions, paralleling"molecular switch", the diminutive G protein in activated state being able to activate the wink dromo-iter to work functions, which is in the resemblance of process during which the Ras genic mutation induces its continous activations.

RhoA是近年来新发现的与多种恶性肿瘤密切相关的癌基因，RhoA属于Rho亚家族，是ras基因超家族的一员，其编码产物RhoA蛋白属于小分子量GTP结合蛋白族，分为GTP结合的活性型和GDP结合的非活性型两类，在正常细胞内以GDP结合的非活性型存在，两种活性形式之间的转换使他们能够发挥一种类似&分子开关&的作用，活性态的小G蛋白能激活下游的信号传导通路，行使其功能。

Do you know, i need you to come back

你知道吗，我需要你回来

Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。