Molecular cloning and tissue distribution of a short form chicken leptin receptor mRNA     

Liu X  Dunn IC  Sharp PJ  Boswell T 《Domestic animal endocrinology》2007,32(3):155-166

In mammals, alternative splicing of the leptin receptor (LEPR) produces several C-terminal truncated isoforms that are believed to play a role in the transport, cellular internalisation and degradation of the hormone leptin. The chicken leptin receptor (chLEPR) is similar to its mammalian counterparts in terms of its intron/exon structure and conserved motifs. However, it is unknown whether the chLEPR also undergoes alternative splicing. To test this, structural analysis of intron 19 of the chLEPR, equivalent to the intron in which alternative splicing occurs in mammals, was combined with 3'-rapid amplification of cDNA ends (3'-RACE) to search for chLEPR splice variants. A 44-amino acid alternative exon 20 was identified that is spliced to generate a short isoform of the chLEPR (chLEPR-SF). Comparative sequence analysis of intron 19 identified two regions that are highly conserved between the chicken and mammals, indicating their possible importance as intronic elements in the regulation of alternative splicing of the LEPR in vertebrates. Tissue expression of the chLEPR-SF was lower and more restricted than that of the chLEPR long isoform. Collectively these data demonstrate that the chLEPR is alternatively spliced to produce at least one short isoform, as is the case in mammals.  相似文献   

二化螟dsRNA降解酶基因的克隆与表达分析     

彭英传  江婷  朱玉麟  张万娜  张晶  韩召军  肖海军 《植物保护学报》2023,50(3):610-619

为解析二化螟Chilo suppressalis体内参与降解双链RNA(double-stranded RNA,dsRNA)的关键核酸酶的功能,克隆二化螟不同的非专一性核酸酶（non-specific nuclease,NUC）基因,并对这些基因进行生物信息学分析和组织定量表达分析,同时对dsRNA降解酶（dsRNA degrading nuclease,dsRNase）活力的组织分布进行研究。结果显示,共克隆获得5个NUC基因,其中有4个编码dsRNase亚家族基因（CsdsRNase1～CsdsRNase4）和1个编码Endonuclease G亚家族基因（CsEndoG）。5个NUC基因的开放阅读框核苷酸序列长度范围为828～1 338 bp,编码275～445个氨基酸残基,其分子量大小为31.68～49.57 kD,预测等电点为5.48～9.42。CsdsRNase1和CsdsRNase2含有信号肽序列,两者相似度极高,且与家蚕Bombyx mori和斜纹夜蛾Spodoptera litura中具有dsRNA降解酶活力的dsRNase同源聚类;CsdsRNase1和CsdsRN...  相似文献   

分子标记技术及其在稻瘟病抗性基因中的应用     

王羡国  何琳  靳学慧  张亚玲 《种子世界》2006,(2):32-34

分子标记技术是近20年发展起来的一种分子生物学技术，具有准确度高、多态性高、表现共显性等特点。根据检测手段的不同，分子标记技术可分为以DNA-DNA杂交为基础、以PCR技术为基础、以DNA杂交和PCR技术相结合为基础和以单核苷酸多态性为基础的4种类型分子标记技术。研究者利用各种分子标记技术已鉴定出30多个水稻抗稻瘟病基因，并进行了基因定位，其中抗稻瘟病基因Pi-ta和Pi-b已被克隆。本文简要介绍了多种分子标记技术及其在抗稻瘟病基因研究中的应用进展。  相似文献   

新城疫病毒La Sota株HN基因的克隆与序列分析     

刘颖  金丽颖  申燕  高冬妮  平文祥  葛菁萍 《中国农学通报》2015,31(14):89-95

旨在从分子生物学角度进一步研究新城疫病毒La Sota株HN基因,探究NDV La Sota HN基因的遗传变异情况。研究以试验室冻存的p NDV-HN为模版,根据Gen Bank中登录的NDV La Sota株序列设计引物扩增HN基因,将其克隆到pMD18-T载体后进行鉴定,对鉴定正确的重组质粒p MD-NDV-HN进行测序分析。应用生物信息学软件对新城疫病毒Lasota株HN基因进行系统进化树分析、氨基酸序列同源性分析、糖基化位点、蛋白结构跨膜区分析及磷酸化位点预测分析。核苷酸序列测定结果表明:HN基因的序列长度为1743bp,该基因的开放阅读框(Open Reading Frame,ORF)总长为1716 bp,编码571个氨基酸。生物信息学表明:试验获得的HN基因具有6个潜在糖基化位点及12个半胱氨酸残基,第5个潜在糖基化位点(508-510 aa)发生缺失及第123位半胱氨酸残基被色氨酸残基取代。La Sota株HN基因编码的蛋白质中有29个丝氨酸、10个酪氨酸和16个苏氨酸可能成为蛋白激酶磷酸化位点。将试验获得的La Sota株HN基因序列和推导的氨基酸序列与新城疫毒株的HN基因相应序列比较后发现,它们的核苷酸序列同源性和氨基酸序列同源性最高均可达到99%,表明HN基因在种属间具有较高的保守性。研究为新城疫病毒的分子病毒学研究奠定了基础。  相似文献   

农杆菌介导遗传转化在水稻基因工程育种中的应用   总被引：5，自引：0，他引：5  

吕彦  王平荣  孙业盈  董春林  陈德西  邓晓建 《分子植物育种》2005,3(4):543-549

随着生物技术的发展，水稻基因工程育种在水稻育种中发挥着越来越重要的作用。在诸多的转基因方法中，农杆菌作为一种天然的植物基因转化系统，以其独特优点而倍受重视。本文概述了农杆菌介导转化水稻的原理，农杆菌介导转化水稻的优点并且介绍了农杆菌菌株、受体材料和培养基成分的不同对农杆菌介导转化水稻的影响。另外对农杆菌介导遗传转化在水稻抗虫基因工程育种、抗病基因工程育种、抗逆基因工程育种、优质基因工程育种、耐除草剂基因工程育种及提高水稻光合效率和延缓衰老方面的研究进展进行了综述。最后对农杆菌介导遗传转化在水稻基因工程育种中存在的问题和发展前景做了简要说明。  相似文献   

炭疽菌附着胞的研究进展   总被引：3，自引：0，他引：3  

王葵娣  王文华  郑服丛 《中国农学通报》2007,23(1):265-265

摘 要：炭疽菌属Colletotrichum Corda 是一类重要的植物病原菌，寄主范围广，对农林生产危害很大。炭疽菌能产生特殊的侵染结构附着胞穿透寄主组织，附着胞的形成对炭疽菌确立侵染非常关键。综述了炭疽菌分生孢子在寄主表面的附着、附着胞的形成、附着胞对环境影响因素的应答和附着胞的穿透。影响炭疽菌附着胞形态发育的因素很多，其中硬质疏水表面在短时间内通常是有利于附着胞的形成。并简要概述了炭疽菌附着胞的基因克隆研究情况。  相似文献   

龙牙百合的植株再生与遗传转化   总被引：19，自引：0，他引：19  

刘菊华  金志强  徐碧玉  郑思乡  刘志敏 《分子植物育种》2003,1(4):465-474

以龙牙百合鳞片叶切块为外植体，通过多种培养基的比较试验，得出MS 2，4-D 2mg／L 6-BA 0．2mg／L是诱导愈伤的最佳培养基，MS 6-BA 1mg／L NAA 0．05mg／L是不定芽诱导及伸长的适宜培养基。MS N_AA2mg／L是生根的最佳培养基。本试验已建立了适用于龙牙百合遗传转化的快速高频再生系统。用携带有几丁质酶基因和β—1、3葡聚糖酶基因的工程菌，通过农杆菌介导法和基因枪转化法转化龙牙百合，经PCR和点杂交检测证明外源基因已经整合到植物染色体中。同时对农杆菌介导法和基因枪法进行比较，发现农杆菌介导法的转化率为16．7％，基因枪法的转化率为50％，因此可能基因枪转化法更适于龙牙百合的遗传转化。  相似文献   

鸡白介素-6(ChIL-6)基因的克隆与序列分析   总被引：7，自引：1，他引：7  

孙建和  陆苹  蒋静 《华中农业大学学报》2003,22(1):1-3

分别用鸡传染性法氏囊病病毒感染和用ConA免疫刺激健康鸡，无菌取处理鸡的脾脏，匀浆后用Tr-izol抽提细胞总RAN，应用随机引物反转录获得cDNA，设计引物，经PCR扩增获得鸡白介素-6（ChIL-6)基因，应用pGEM-T载体克隆该基因，经测序，表明其与Gene Bank中公布的唯一的一个ChIL-6基因为同源基因。  相似文献   

酵母基因表达调控关系的构建及其统计特性分析   总被引：1，自引：0，他引：1  

吉冬梅 《安徽农业科学》2009,37(10):4360-4362

[目的]从复杂网络的角度研究酵母细胞周期调控网络的统计特性。[方法]首先对酵母的细胞周期基因表达数据采用皮尔逊相关性度量,计算基因间的关联矩阵,即建立简单的基因调控网络;同时为了能够得到在宏观角度对大规模的基因调控有一个全貌的了解,进一步分析所建立调控网络的统计特性,通过计算所建立网络的统计特性：即计算网络的聚类系数、平均路径长度和网络的度分布。[结论]发现所建立的网络具有无标度和小世界特性,即所建立的酵母基因调控网络属于复杂网络,这为进一步深入研究基因调控网络的统计学和动力学特性打下了基础。  相似文献   

CD147 monoclonal antibody-mediated nanoparticles for gene therapy to target lung cancer cells     

WU Fei-long  KONG Qing-lei  CAI Song-wang  YE Zhi-qiang 《园艺学报》2016,32(9):1562-1567

AIM: In this study, CD147 antibody was used to carry out targeted modification of nanoparticles for protein kinase Cε (PKCε)-siRNA gene therapy to target lung cancer cells. The inhibitory effects of the nanoparticles on the proliferation and invasion of the lung cancer cells were observed. METHODS: The magnetic nanoparticles targeting CD147 protein were assembled as gene vector. The expression of CD147 in the lung cancer cells was observed under laser scanning confocal microscope. The cells were divided into CP group, CN group and LP group as the experimental groups. Targeted nanoparticles were used as CA group. Non-transfected cells were used as control group. The cell transfection was carried out with 250 ng plasmids/well in 6-well plate. The effect of nanocontrast agent on the cell endocytosis was observed under laser scanning confocal microscope. The mRNA expression of PKCε was detected by RT-qPCR. The protein expression of Ki67, MMP3, PKCε, Wnt1 and GAPDH was determined by Western blot. The cell proliferation ability was detected with colony formation assay. The cell invasion ability was detected by Transwell method. RESULTS: The expression of CD147 protein in the human lung cancer A549 cells was confirmed by immunofluorescence staining. The endocytosis of siRNA into the A549 cells in CP group was observed with the highest efficiency as compared with CN group and LP group. The relative mRNA expression of PKCε in the A549 cells of CP group, CN group, LP group and CA group were (9.76±0.18)%, (98.51±0.32)%, (99.17±0.16)% and (99.68±0.11)%, respectively. The difference between CP group and control group was statistically significant (P<0.05). No significant difference among CN group, LP group and control group was observed. The protein expression of PKCε, Ki-67, MMP3 and Wnt1 in CP group was significantly reduced, and the protein expression levels among CN group, LP group and control group had no significant difference. The colony number in CP group was significantly smaller than that in control group (P<0.05). The effective colony numbers in CN group, LP group and CA group had no significant difference as compared with control group. The number of the invading cells in CP group was significantly less than that in control group (P<0.05). The numbers of the invading cells in CN group, LP group and CA group had no significant difference as compared with control group. CONCLUSION: Nanogene vector targeting CD147 can carry PKCε-siRNA to conduct gene therapy efficiently on the lung cancer cells to achieve effective inhibitory effects on the proliferation and invasion of the lung cancer cells.  相似文献