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Rapid alkalinization factors were polypeptide signals in the plant, belonging to a large gene family. The activity of RALF could induce extra-cellular pH increasing rapidly. Many studies on RALFs were in Solanum, including the structure and maturation, the gene expression, receptors and functions and so on.

快速碱化因子类基因属于植物多肽类信号分子,是一类大的家族基因,它的活动引起胞外pH值快速升高,有关RALF类基因的研究在几种茄科植物中开展的较多,包括RALF蛋白信号分子的结构及加工、基因表达、受体研究以及功能等方面。

Objective: The purposes of the present study are as follows: 1 To study the effect of direct contact with VSMC on transdifferentiation of MSC; 2 To construct MSC of Dual-stable expression of AT2R gene medicated by tetracycline regulatable system.; 3 To investigate the effects of AT2R gene medicated by tetracycline regulatable system on neointimal hyperplasia in rat carotid arteries after balloon angioplasty.

本实验体外观察MSC与VSMC在直接接触培养下的分化情况,并以MSC为载体,通过体外细胞基因转染及筛选,获得双重稳定受到强力霉素调控表达AT2R基因的MSC,将此受到调控表达的AT2R基因导入大鼠颈动脉球囊损伤动物模型中,观察MSC细胞移植实现AT2R基因在体可调控表达在新生内膜中形成的作用及潜在意义。

In order to improve the resistance to bacterial blight of high quality rice male sterile line Jinshan A , in this study, the Jinshan B and IRBB21 that carried gene Xa21 were used as parents to make backcrosses, and both foreground selection and background selection were carried out by means of marker assisted selection in the backcrosses.

本研究以IRBB21为Xa21基因的供体亲本,以金山B为轮回亲本进行回交,并应用分子标记辅助选择技术对回交各世代同时进行前景选择和背景选择,得到如下结果:(1)利用Xa21基因内的STS标记MXA21对各回交世代进行前景选择,得到了44个带有Xa21基因的BC_1单株和40个带有Xa21基因的BC_2单株。

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。

These constructs, together with the construct containing the GUS gene expression cassette under the control of the Hsp70 promoter, were utilized in the co-bombardment experiments to identify viral cistrons that could activate the transcription directed by the Hsp70 promoter. We found that the P1, K2NIa and CP cistrons could independently activate the Hsp70 promoter.

用这些瞬时表达载体与含有Hsp70基因启动子指导下的GUS基因的表达载体共轰击豌豆叶片,发现P1、K2NIa和CP能诱导Hsp70基因启动子的转录活性,P1最强,K2NIa次之,而CP最弱,同时还发现P1基因的功能区域在127碱基以后。

The possibility of researching gene mutation with the technique of gene chip was discussed. A new imagination to randomly determine DNA sequence in different living beings with the technique of gene chip was advanced. In this way, whether the encoding sequence gene in DNA of eukaryon organism has some regularity was studied.

进而提出新设想——采用基因芯片技术对不同生物群体随机抽样进行DNA测序,从而探讨在真核生物DNA分子链上编码序列遗传基因的变化是否有规则而非随机的,非编码序列遗传基因是否为生物进化过程中曾经有表达能力的结构基因,并分析可能获得的结果。

Total RNAs from KAx-3 cells and AK127 cells(developed for 14h) were isolated. After the reverse transcription and PCR reaction, two distinct differential fragments were acquired., fragment A was from KAx-3 cells and fragment C was from AK127 cells. After retriving and reamplifying the differentially expressed fragments, white-blue plaqueselection, the fragments were purified. Northern blot proved that fragment A was from KAx-3 cells and fragment C was from AK127 cells. The results of sequencing and researching for NCBI database have been showed: part sequence of fragment A shows 91% similarity to the gene encoding DhkA, 92% similarity to the gene encoding DhkF, 91% similarity to the gene encoding STATc, 97% similarity to the homoeobox gene encoding protein. These genes play important part in controlling cell differentiation and cell proportion in Dictyostelium discoideum.

本研究通过提取盘基网柄菌发育14小时的野生型KAx-3细胞和突变型AK127细胞的总RNA,运用mRNA差异显示法分离出了两条明显的差异表达片段,其中片段A来自野生型KAX-3细胞,片段C来自突变型Ak127细胞;并通过凝胶回收差异片段、对差异片段进行再次PCR、蓝白斑筛选克隆、提取质粒、酶切电泳纯化差异片段;接着进行Northern杂交的结果表明,片段A只与野生型KAx-3细胞的总RNA有杂交信号,片段C只与突变型AK127细胞的总RNA有杂交信号,这就排除了差异片段假阳性的可能;最后通过测序,搜索NCBI BLAST数据库发现:片段A的小部分序列与编码组氨酸激酶DhkA基因中一段序列的相似性高达91%,与编码组氨酸激酶DhkF基因中的一段序列相似性高达92%,与编码STATc蛋白基因的一段序列相似性达91%,以及与编码同源框蛋白的基因中的一段序列相似性达97%,这些基因在盘基网柄菌细胞分化和细胞比例调控过程中起着相当重要的作用,这些数据进一步说明了突变细胞不能完成发育的原因。

It may help explain how some of the world's most glamorous women are able to maintain their slimline figures so easily. Superslim models Naomi Campbell and Kate Moss both have skinny mothers.

研究人员表示,体内携带这种基因的女性比没有携带此类基因的女性更瘦一些,而且携带&瘦体&基因的女性的孩子更容易从母亲那里遗传到这种基因。

Xanthus. The plasmid vector pZCY11 made it more convenient for the study on functions and the expressions of target gene in M. xanthus.

构建的质粒载体pZCY11不仅能够对目的基因进行功能的分析,而且能够同时通过报告基因分析基因的表达情况,可简化粘球菌中基因功能及表达情况的研究。

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