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1.
【目的】分析苏博美利奴羊不同组织中Hoxc13基因mRNA的表达量差异,为分子标记辅助育种以及,研究细毛羊主要经济性状,奠定理论基础。【方法】使用RT-PCR技术检测Hoxc13基因在苏博美利奴羊65和135 d皮肤、肌肉和不同内脏组织(心脏、脾脏、肝脏、肺脏、肾脏)中mRNA的表达量,并采用相对定量法对其表达量进行统计分析。【结果】在苏博美利奴羊皮肤、肌肉和不同内脏组织中均有Hoxc13基因的表达,并且在皮肤中的表达量显著高于其它组织(P<0.05),毛囊的形成和毛发生长与Hoxc13基因密切相关。【结论】建立了胎龄分别在65和135 d的苏博美利奴羊,其不同组织中适于定量检测的Hoxc13基因表达量的RT-PCR方法,从分子水平探讨了苏博美利奴羊的组织器官与Hoxc13基因表达之间的内在联系。  相似文献   

2.
为研究光控增绒和埋置褪黑激素促进绒山羊在非产绒季节(5—8月份)长绒的皮肤毛囊差异表达基因之间的互作关系,并筛选出皮肤毛囊发育相关的重要功能基因,以Agilent绵羊的全基因组表达谱芯片杂交获得的光控增绒羊皮肤差异表达的99个基因及埋置褪黑激素绒山羊皮肤差异表达的83个基因作为研究对象,结合NCBI数据库,利用RStudio程序包和Cytoscape 3.3.0构建差异基因网络并分析。光控增绒试验组差异基因构建了1个主网络和4个小网络;对照组差异基因构建了1个主网络和5个小网络。与表达量结合分析基因网络发现,K25、KAP16-2基因上调促进ISG17、RSRP1、GIMAP1和CYTIP基因的下调,间接促进TNFAIP6基因的下调;CYP17A1、V15和K2-12(K85)基因的上调促进ddit3基因的下调;KAP16-3基因促进CYP17A1基因的同时抑制CYP1A1基因。埋置褪黑激素试验组差异基因构建了2个主网络和7个小网络,对照组差异基因构建了3个主网络和1个小网络。与表达量结合分析基因网络发现,CTNNB1基因的上调促进COL3A1基因的下调;FLT-1基因的上调促进CGA基因的上调和COL1A2基因的下调;PAX6基因的上调促进DOK4基因的上调和LOC101110099基因的下调;P450基因的上调促进BLG基因的下调。光控增绒羊皮肤中筛选出来的基因中角蛋白、角蛋白关联蛋白、CYP17A1、SP-D等23个显著上调基因和CYP1A1、COL6A5、LOC100101238、ddit3等13个显著下调基因对光控增绒羊皮肤毛囊提前进入兴盛期有重要作用;埋置褪黑激素的绒山羊皮肤中筛选出来的基因中CTNNB1、PAX6、DOK4、LOC105602529等13个显著上调基因和酪蛋白家族、胶原蛋白类、LOC101110099等12个显著下调基因之间的信号传导增强,从而促进绒山羊绒毛进入兴盛期。  相似文献   

3.
WUSCHEL-related homeobox基因编码保守的转录因子家族在植物发育中起着非常重要的作用。本文从茄子中克隆了SmWOX13基因,该基因cDNA序列全长1 233 bp,包含834 bp编码序列。推导氨基酸序列用于分析该基因的保守性及生成3D结构。扩增获得的基因组序列被用于该基因结构的分析。SmWOX13与GFP融合蛋白的亚细胞定位结果表明该蛋白被定位于细胞核。系统发生树显示茄子SmWOX13基因与番茄WOX8和拟南芥WOX13属于同一个分支。综上结果可表明茄子SmWOX13基因可能与之前报道的番茄和拟南芥WOX基因类似,参与了果实形状的控制。  相似文献   

4.
WUSCHEL-related homeobox基因编码保守的转录因子家族在植物发育中起着非常重要的作用。本文从茄子中克隆了SmWOX13基因,该基因cDNA序列全长1 233bp,包含834bp编码序列。推导氨基酸序列用于分析该基因的保守性及生成3D结构。扩增获得的基因组序列被用于该基因结构的分析。SmWOX13与GFP融合蛋白的亚细胞定位结果表明该蛋白被定位于细胞核。系统发生树显示茄子SmWOX13基因与番茄WOX8和拟南芥WOX13属于同一个分支。综上结果可表明茄子SmWOX13基因可能与之前报道的番茄和拟南芥WOX基因类似,参与了果实形状的控制。  相似文献   

5.
为挖掘脊椎动物胚胎发育过程中皮肤调控模式的改变,以鸡胚作为模型,利用来源于NCBI数据库的SRA数据子库中胚胎发育第6~21天的皮肤组织RNA-Seq数据进行聚类分析,在基因共表达网络、信号通路、蛋白互作网路3个层面基于网络拓扑属性逐步挖掘核心基因集,并分析核心基因集在不同发育阶段的互作网络变化。结果表明:1)鸡胚皮肤发育过程在基因谱水平上可分成5个阶段,分别为6~9 d、10~12 d、13~14 d、15~17 d和18~21 d;2)功能富集分析表明发育早期的皮肤分化程度低、可塑性强,中期是皮肤毛囊发育关键期,最后4 d为皮肤细胞外基质建立期,分子发育过程存在严格的顺序模式;3)在基因调控网络层面深度挖掘并筛选鸡胚皮肤发育的分子调控过程。综上,本研究根据网路拓扑属性结合蛋白互作数据库提出参与不同发育模块的关键枢纽基因,为转录组测序数据的分析及脊椎动物皮肤发育过程的分子机制研究提供一些新的视角和分析方法。  相似文献   

6.
利用RACE-PCR技术获得了牙鲆FREP1(fibrinogen-related protein)基因1 131 bp cDNA全长序列,其中开放阅读框786 bp,所编码的蛋白C端含有纤维蛋白样结构域标签(WWYSRCGSAGLNG).RNA整体原位杂交检测发现只在孵化后2d和5 d(2 dph、5 dph)仔鱼的肠道中有FREP1基因表达,而在9 dph和13 dph仔鱼胸鳍、肠道、鳃和皮肤中均有表达.荧光定量PCR检测显示该基因主要在成鱼的肠、鳃、皮肤和鳍条上表达,而脾脏、心脏、肾脏、肝脏和肌肉表达量很低或没有表达.鳍条和皮肤较其他组织均有极显著差异(P<0.01),鳃和肠相比其他组织具有显著差异(P<0.05).由于该基因主要在鳍条、皮肤、肠和鳃中表达,提示FREP1基因可能和牙鲆先天性免疫相关.研究亮点:FREP在鱼类免疫方面的报道很少,本研究所克隆的牙鲆FREP1,不管是牙鲆仔鱼还是成鱼,其仅在与外界直接接触的皮肤、鳃、肠道等组织中表达,提示其与牙鲆的先天免疫相关.  相似文献   

7.
利用RACE-PCR技术获得了牙鲆FREP1(fibrinogen related protein)基因1 131 bp cDNA全长序列,其中开放阅读框786 bp ,所编码的蛋白C 端含有纤维蛋白样结构域标签(WWYSRCGSAGLNG)。RNA整体原位杂交检测发现只在孵化后2 d和5 d(2 dph、5 dph)仔鱼的肠道中有FREP1基因表达,而在9 dph和13 dph仔鱼胸鳍、肠道、鳃和皮肤中均有表达。荧光定量PCR检测显示该基因主要在成鱼的肠、鳃、皮肤和鳍条上表达,而脾脏、心脏、肾脏、肝脏和肌肉表达量很低或没有表达。鳍条和皮肤较其他组织均有极显著差异(P<0.01),鳃和肠相比其他组织具有显著差异(P<0.05)。由于该基因主要在鳍条、皮肤、肠和鳃中表达,提示FREP1基因可能和牙鲆先天性免疫相关。  相似文献   

8.
【目的】研究角蛋白相关蛋白9.2基因(KAP9.2)和Hoxc13基因在羊绒生长不同阶段皮肤组织中的表达量对产绒量的影响,为羊绒生长的调控机理研究奠定基础。【方法】以陕北白绒山羊为研究对象,以奶山羊作为对照,利用qRT-PCR方法,检测羊绒生长期和休止期高产、低产绒量陕北白绒山羊皮肤组织中KAP9.2和Hoxc13基因的表达量。【结果】 对于KAP9.2基因,在羊绒生长期高产绒量陕北白绒山羊的相对表达量极显著低于低产绒量陕北白绒山羊(P<0.01),而在羊绒休止期高产绒量陕北白绒山羊的相对表达量却显著高于低产绒量陕北白绒山羊和奶山羊(P<0.05);对于Hoxc13基因,在羊绒休止期高产和低产绒量陕北白绒山羊的相对表达量极显著高于奶山羊(P<0.01)。高产和低产绒量陕北白绒山羊皮肤组织中KAP9.2和Hoxc13基因在生长期、休止期平均相对表达量的变化趋势相似,均表现为生长期极显著低于休止期(P<0.01)。【结论】KAP9.2对绒毛生长有抑制作用;Hoxc13对KAP9.2基因的表达可能具有调控作用。  相似文献   

9.
尖裸鲤实时荧光定量PCR内参基因的筛选   总被引:1,自引:0,他引:1  
为筛选能够稳定表达的内参基因用于尖裸鲤Oxygymnocypris stewarti实时荧光定量PCR分析,以尖裸鲤血液、心脏、肝、脾、鳃、头肾、中肾、后肾、前肠、中肠、后肠、性腺、眼、垂体、脑、红肌、白肌和皮肤18个组织为材料,应用实时荧光定量PCR技术比较了GAPDH(3-磷酸甘油醛脱氢酶)、HPRT1(次黄嘌呤鸟嘌呤磷酸核糖转移酶1)、RPL13(核糖体蛋白L13)、RPL19(核糖体蛋白L19)、RPL13a(核糖体蛋白L13a)、SDHA(琥珀酸脱氢酶亚基A)和ACTB(β-肌动蛋白)7个候选内参基因的表达情况,并采用GeNorm、NormFinder和BestKeeper软件对7个候选内参基因的稳定性进行分析。结果表明:GeNorm分析显示,7个候选内参基因的平均表达稳定值(M)依次为RPL13=RPL13aRPL19GAPDHACTBSDHAHPRT1,其中RPL13和RPL13a的M值均最小(0.62);NormFinder分析显示,7个候选内参基因的M值依次为RPL13RPL13aRPL19GAPDHACTBSDHAHPRT1,其中RPL13的M值最小(0.30);BestKeeper分析显示,7个候选内参基因的标准差(S.D.)依次为RPL13RPL19GAPDHHPRT1RPL13aSDHAACTB,变异系数(CV)值依次为RPL13RPL19HPRT1GAPDHSDHARPL13aACTB,其中RPL13的标准差(0.84)和变异系数(4.54%)均最小;7个候选内参基因在尖裸鲤不同组织中的表达量存在差异。研究表明,7个内参基因中RPL13的表达较为稳定,可作为尖裸鲤实时荧光定量PCR分析的适宜内参基因,本研究结果可为尖裸鲤遗传学及分子生物学等研究提供基础数据。  相似文献   

10.
采用目前国际上最为通用的cDNA文库构建方法,构建了银鲳皮肤和肌肉组织的cDNA文库,通过一定规模的序列测定,并借助生物信息学技术,分析了164个序列,得到了105个不同的基因(或表达序列标签),在105个不同的基因中,68.6%基因没有冗余,1倍、2倍、3倍和4倍冗余的分别占23.8%,3.8%, 2.8%和1%.皮肤和肌肉组织表达基因的组成中,除了21个没有同源序列的未知基因外,可初步归为代谢相关的酶(38个)、DNA有关的酶(7个)、结构蛋白(4个)、膜蛋白(13个)、线粒体蛋白(1个)、转录因子(9个)、信号传导相关的蛋白(10个)和未知蛋白(2个)等8大类,其中,转录因子、膜蛋白,和DNA有关的酶的基因冗余程度相对较高.这些研究结果,为积累银鲳遗传方面的基础信息,了解银鲳皮肤和肌肉组织的基因表达提供了较好的研究基础.  相似文献   

11.
Human primary skin fibroblasts trisomic for chromosome 13, 18, or 21 and diploid human skin fibroblasts were induced for an antiviral response with human interferon. The cells that were trisomnic for chromosome 21 were three to seven times more sensitive to protection by human interferon than the normal diploid or trisomic 18 or 13 fibroblasts. The differential response in trisomnic 21 cells is consistent with the known assignment of the human antiviral gene to chromosome 21.  相似文献   

12.
Seasonal hair follicle activity and fibre growth in some Cashmere-bearing goats (Caprus hircus) is a cyclic process that is well characterized morphologically but understood incompletely at the molecular level. As an initial step in discovering regulators in hair-follicle activity and cycling, we used qPCR to investigate 19 genes expression in Cashmere goat side skin from 12 mon. Many of these genes may be associated with the hair follicle development-relevant genes (HFDRGs) in the literature. Here we show that Hoxc13/β-catenin gene associated with the follicle activity. In addition, Hoxc13 was found to be expressed with an drastic increase between July and November for melatonin treatments. To further investigate the role of Hoxc13 on HFDRGs, fibroblasts and keratinocytes from Cashmere goat skin were transfected with p-ECFPHoxc13. The result suggested that overexpression of Hoxc13 gene decreased HFDRGs with negative role for hair follicle development and increase HFDRGs with positive role for hair follicle development in vitro. These findings provide data on the Hoxc13 expression profile of normal Cashmere goat skin and Cashmere goat skin with melatonin treatment, and demonstrate hair-follicle-activity dependent regulation of Hoxc13 expression.  相似文献   

13.
Hoxc13 has an important role in controlling hair formation. In this study, we examine the Hoxc13 RNA expression pattern of skin during embryo development. The result indicated that changes of the Hoxe13 gene expression and thickness of skin have a similar trend during hair follicle morphogenesis. In interpreting these results, we investigated whether the regulation motifs is in Hoxc13 intron, which is a 5.4 kb fragment. To blast with other mammals, we found a very conservative region in all mammal animals and two regions in livestock, such as cow, sheep, horse, dog, and so on, which are not in other Hox genes. We have examined putative pre-miRNA in this region, providing an entry point for elucidating currently unknown mechanisms that are required for regulating quantitative levels of Hoxc13 gene expression.  相似文献   

14.
用Ⅰ型、Ⅳ型胶原酶消化分离兔皮肤细胞,Ⅰ型胶原酶消化皮肤块获得的细胞数显著高于Ⅳ型胶原酶。用组织块直接培养法,也可以得到皮肤的原代培养细胞。2.5g/L胰酶消化胎儿组织,可获得足够原代培养的细胞量。结合使用酶消化法和反复贴壁法,经2~3代后,可获得纯化的成纤维细胞。分离纯化的胎儿和皮肤成纤维细胞的生长曲线都正常且相似。胎儿和皮肤成纤维细胞可分别传至第13代与第11代,传代后染色体数目不变。  相似文献   

15.
Alterations in NMRI mouse skin induced by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate in "stage I of tumor promotion" are slowly reversible, and this reversibility has a half-time of 10 to 12 weeks. The tumor response observed in the course of an initiation-promotion experiment in vivo is independent of whether stage I of promotion occurs before or after initiation. Since the time interval between treatment with the promoter, and subsequent initiation can be extended up to at least 6 weeks, an enhancement of initiation because of promoter-induced cellular DNA synthesis seems to be unlikely. This result may be inconsistent with the two-stage model of tumor promotion because it indicates that in skin the existence of initiated cells is not required for the induction of cellular alterations that are essential for the stage of skin tumorigenesis called stage I of promotion.  相似文献   

16.
Although recent experimental studies have suggested that the interactions among the pigment cells play a key role in the skin pattern formation, details of the mechanism remain largely unknown. By using an in vitro cell culture system, we have detected interactions between the two pigment cell types, melanophores and xanthophores, in the zebrafish skin. During primary culture, the melanophore membrane transiently depolarizes when contacted with the dendrites of a xanthophore. This depolarization triggers melanophore migration to avoid further contact with the xanthophores. Cell depolarization and repulsive movement were not observed in pigment cells with the jaguar mutant, which shows defective segregation of melanophores and xanthophores. The depolarization-repulsion of wild-type pigment cells may explain the pigment cell behaviors generating the stripe pattern of zebrafish.  相似文献   

17.
目的观察系统性红斑狼疮(SLE)患者皮损中IL-36β、IL-36R表达。方法应用免疫组化技术检测13例SLE患者皮损和8例对照者皮肤中IL-36、IL-36R分布及表达水平。结果 SLE患者皮损中表皮角质形成细胞、真皮浅层浸润的淋巴细胞核表达IL-36、IL-36R水平较对照组高(P〈0.01)。结论 SLE患者皮损中存在IL-36和IL-36R高表达,可能参与SLE发病。  相似文献   

18.
19.
Levin BR  Antia R 《Science (New York, N.Y.)》2001,292(5519):1112-1115
To pathogenic microparasites (viruses, bacteria, protozoa, or fungi), we and other mammals (living organisms at large) are little more than soft, thin-walled flasks of culture media. Almost every time we eat, brush our teeth, scrape our skin, have sex, get bitten by insects, and inhale, we are confronted with populations of microbes that are capable of colonizing the mucosa lining our orifices and alimentary tract and proliferating in fluids and cells within us. Nevertheless, we rarely get sick, much less succumb to these infections. The massive numbers of bacteria and other micro- and not-so-micro organisms that abound and replicate in our alimentary tract and cover our skin and the mucosa lining our orifices normally maintain their communities in seemingly peaceful coexistence with the somatic cells that define us. Why don't these microbes invade and proliferate in the culture media within the soft, thin-walled flask that envelops us? Why don't they cause disease and lead to our rapid demise?  相似文献   

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