共查询到19条相似文献，搜索用时 203 毫秒
1.
促卵泡素(FSH)是动物脑垂体分泌作用于卵巢卵泡生长、发育、分化、成熟和排卵的重要繁殖激素.由于是生物大分子,不能透过细胞膜,FSH必须与靶细胞膜上的促卵泡素受体(FSHR)结合,经过受体介导将信息传递到靶细胞内,才能发挥其生物学功能.目前国内外已对牛、山羊等多种动物的FSHR基因的Cdna序列进行了克隆测序,但是关于水牛FSHR基因5'端侧翼序列还未见报道.本研究对水牛FSHR基因5'端序列进行克隆和序列分析,其目的是寻找水牛FSH受体基因的SNP突变位点,从而为以后进行FSH受体基因的多态性分析和探索该受体基因在水牛繁殖中的可能作用打下基础. 相似文献
2.
促卵泡素(FSH)是动物脑垂体分泌作用于卵巢卵泡生长、发育、分化、成熟和排卵的重要繁殖激素。由于是生物大分子,不能透过细胞膜,FSH必须与靶细胞膜上的促卵泡素受体(FSHR)结合,经过受体介导将信息传递到靶细胞内,才能发挥其生物学功能。目前国内外已对牛、山羊等多种动物的FSHR基因的cDNA序列进行了克隆测序,但是关于水牛FSHR基因5′端侧翼序列还未见报道。本研究对水牛FSHR基因5′端序列进行克隆和序列分析,其目的是寻找水牛FSH受体基因的SNP突变位点,从而为以后进行FSH受体基因的多态性分析和探索该受体基因在水牛繁殖中的可能作用打下基础。由水牛血液中提取DNA,与根据牛FSH受体基因序列设计的特定引物进行PCR反应,以合成FSHR 5′端侧翼调控区,扩增的该片断经分离、回收和纯化后再插入到pMD18-T质粒中,筛选阳性克隆,将插入了FSHR 5′端侧翼端片断的质粒纯化后进行序列测定和分析,弄清核苷酸序列和假定的氨基酸顺序并通过GeneBank进行同源性分析。结果表明,PCR扩增获得的片段为939 bp,与中国西门塔尔牛该段序列相比,同源性为97.44%,在-720 bp处的1个碱基缺失;与山羊的FSHR基因比较,同源性为94.78%,在-625~-619 bp处有7个碱基的插入;在此激素反应元件3′端一段序列(-530~-526 bp)水牛和山羊的相同均为TGACC,而西门塔尔牛为CGACC,但三者5′端一段序列(-679~-675 bp)没有突变并且相同,均为GGTCA;与人和大鼠FSHR基因不同的是,在牛、水牛和羊的FSHR基因启动子区检索出了TATA盒基元和多个类CAAT盒序列,且在水牛、山羊和牛之间没有碱基差异。因此,对水牛FSHR基因转录启动和表达调控值得进一步研究。 相似文献
3.
采用RT-PCR方法从水牛外周血白细胞总RNA中扩增出髓样分化因子88（mydoid differentiation factor88,MyD88）cDNA序列,PCR产物分离纯化后,与pMD20-T载体连接,重组质粒经PCR、酶切鉴定后测序,并进行生物信息学分析;构建pET28a-MyD88表达载体,并将其转化至E.coli BL21（DE3）,经IPTG诱导表达后,进行SDS-PAGE、镍柱亲和层析纯化和Western blotting分析。结果显示,克隆到的水牛MyD88cDNA全长为1189bp,含有1个891bp的开放阅读框,编码296个氨基酸,理论等电点为5.65。经IPTG诱导表达后,得到一个带His·Tag的约39kD的重组融合蛋白。用抗His单克隆抗体进行Western blotting,得到1条约39kD特异性抗体结合带,表明水牛MyD88原核表达载体成功构建并表达。本研究为进一步开展水牛MyD88的结构功能分析奠定了基础。 相似文献
4.
从广西巴马小型猪的卵巢中提取总RNA，并以其为模板，应用逆转录一聚合酶链式反应(RT—PCR)，在合成的特异性引物引导下，扩增得到巴马小型猪卵泡抑素(Follistatin) cDNA的全序列，长度为1035bp。该扩增片段经纯化后，连接到pMD18-T载体上扩增。经序列分析结果表明，本研究中克隆的猪卵泡抑素cDNA序列与GeneBank中已报道的家猪卵泡抑素同源性高达99．4％，与奶牛、家鼠、挪威鼠和马等其他物种的卵泡抑素cDNA序列间同源性也大于89％。 相似文献
5.
本研究克隆了广西沼泽型水牛γ-干扰素(interferon-γ,IFN-γ)基因,并构建了水牛IFN-γ原核表达质粒。从健康沼泽型水牛静脉无菌采血,分离外周血单个核细胞(Peripheral blood mononuclear cells,PBMCs),用刀豆素A(Concanavalin A,ConA)诱导培养13 h后,提取细胞总RNA,用水牛IFN-γ基因特异性引物通过RT-PCR扩增水牛IFN-γ成熟肽编码区cDNA序列,将其克隆到pMD18-T载体中。RT-PCR产物电泳可见约457 bp大小目的片段。经过限制性酶切分析,测序证实克隆得到的基因序列正确。将IFN-γ成熟肽编码区基因片段切下亚克隆到表达载体pET-32a 中,构建成重组表达质粒pET-mIFN-γ。PCR、双酶切电泳和序列测定结果均证实已插入约457 bp的IFN-γ基因片段。经异丙基硫代-β-D-半乳糖苷(IPTG)诱导,水牛IFN-γ基因在大肠杆菌中获得了高效的表达,融合蛋白的分子量为35 ku,表达量占菌体总蛋白的43.6%。 相似文献
6.
本文采用RT-PCR方法,从齿肋赤藓（Syntrichia caninervis）总RNA中获得ALDH21基因cDNA序列,连接到pMD19-T载体上并转化E.coli DH5α,阳性克隆经PCR鉴定后测序,将测序结果与GenBank中山墙藓（Tortula ruralis）和小立碗藓（Physcomitrella patens）相关基因序列进行同源性比对。结果表明,实验成功克隆了S.caninervis的ALDH21基因的cDNA序列（GenBank登录号：GQ245973）,为一个完整的ORF,其长度为1452bp。该序列与T.ruralis和P.patens的cDNA序列同源性分别为98%和78%,推导的氨基酸同源性分别为97%和87%。生物信息学分析表明该蛋白质分子量为52.98kD,等电点pI为5.96,编码483个氨基酸,具备ALDH21蛋白家族特征;半定量RT-PCR结果表明：ALDH21在干旱胁迫状态时的表达量显著高于水合状态,说明ALDH21基因可能参与干旱胁迫应答。本文为进一步研究齿肋赤藓ALDH21基因的抗旱机理提供理论依据。 相似文献
7.
Krtippel样因子4依脾）在调控细胞增殖、分化和胚胎发育中有重要作用,也是生产诱导多能干细胞（iPSC）重要的转录因子之一。本研究对水牛K牌进行克隆和表达研究,为生产水牛iPSC和研究其在体细胞重编程中的机理奠定基础。采用RT—PCR克隆并分析水牛K牌编码区序YIJ（CDS）;mRNA水平和蛋白水平检测K牌在水牛胎儿成纤维细胞（BFF）和胃上皮细胞（BSEC）中的表达情况;构建表达水牛K孵的逆转录病毒载体（pMX—Klf4）、病毒感染BFF、免疫荧光技术分析外源K脾在BFF中的表达情况。结果表明：水牛CDS全长1434bp,氨基酸序列与牛、猪、人和鼠相应氨基酸序列的相似性分别为99％、97％、92％$N92％,蛋白结构保守;内源Klf4在BFF和BSEC中都有表达,pMX介导的外源K脾能在BFF中表达。 相似文献
8.
以广西巴马小型猪的脂肪组织总RNA为模板,通过RT-PCR扩增出巴马小型猪的leptin基因成熟肽序列,经纯化回收后连接到pMD18-T载体上,经鉴定后测序。研究结果表明,本试验克隆到leptin基因成熟肽cDNA序列,其长度为441bp,与GenBank中报道的猪leptin成熟肽cDNA序列同源性高达99．06％,与奶牛、家鼠、马等物种的成熟肽cDNA序列间同源性可达到84．5％以上,证明leptin基因具有较高的保守性。同时发现,与普通猪相比,巴马小型猪leptin基因成熟肽cDNA序列有四处存在碱基突变,均为元义突变。 相似文献
9.
松墨天牛（Monlchamus alternatus Hope）是林业重大害虫。本研究运用SMART技术构建了松墨天牛幼虫cDNA文库,库容量为1.09×10^6,文库滴度为5.45×10^6 pfu/mL,重组率为97.8%,平均插入片段大小约为1 000 bp,构建的文库质量较好。随机选取1 022个克隆进行测序,获得了1 012条表达序列标签（ESTs）,序列在GenBank中登录号为JZ143720～JZ144731。EST序列经过聚类拼接得到411条非重复单一序列,其中216条序列有功能注释。在216条有功能注释的序列中,有181条（83.80%）参与＂分子功能＂,122条（56.48%）参与＂细胞成分＂,168条参与＂分子生物学过程＂（77.78%）。研究结果为克隆和分析松墨天牛新的功能基因奠定了基础。 相似文献
10.
玉兰肌动蛋白基因的克隆与序列分析 总被引：11，自引：0，他引：11
本研究选取玉兰（Magnolia denudata Desr.)为材料，提取总RNA，利用设计的肌动蛋白编码区两端的特异性引物和5'RACE试剂盒，扩增出肌动蛋白编码的cDNA基因。以豌豆卷须肌动蛋白基因作探针，通过Southern杂交进行检测，证明同源性很高。将所获得序列克隆到pGEM-T载体上进行测序，测序结果在GenBank中进行查询，并与其他高等植物肌动蛋白基因序列进行了分析。 相似文献
11.
Organic matter (OM) is the most critical factor in controlling the sorption-desorption of SMZ in soil, however, few studies have explored the effects of OM removal on these important behaviors among different soils. Batch experiments were conducted to investigate the sorption and desorption characteristics of SMZ in three different soils: fluvo-aquic soil (FS), paddy soil (PS), and red soil (RS). The SMZ sorption in the evaluated soils was dominated by physisorption. The SMZ sorption capacities of FS and PS, which had a relatively higher OM content than RS, were higher than that of RS. The SMZ sorption in FS was dominated by linear partitioning. In contrast, the SMZ sorption in PS and RS was mainly nonlinear surface adsorption. After OM removal, the SMZ sorption capacity was significantly reduced in FS but increased in PS and RS. Furthermore, OM removal restrained the sorption intensity of SMZ in soils. Relatively higher OM and clay contents inhibited the SMZ desorption in FS and PS. The strong negative desorption hysteresis of SMZ in the three soils indicated that SMZ was able to move into the soil solution, thereby posing a risk to humans. Taken together, the findings of this study showed that OM indeed plays an important role during SMZ sorption-desorption in soil. 相似文献
12.
为建立表达标记基因的重组猪繁殖与呼吸综合征病毒(PRRSV)反向遗传操作平台,本研究在PRRSV HuN4-F112疫苗株感染性分子克隆的基础上,采用突变PCR技术将新城疫病毒(NDV)NP蛋白末端优势抗原区插入PRRSV NSP2蛋白的复制非必须区,经基因片段的克隆、拼接,构建了含有外源标记基因的感染性的PRRSV cDNA克隆psk-HuN4-F112-Δ52+NP49。用限制性内切酶SwaΙ将psk-HuN4-F112-Δ52+NP49线性化后通过细胞外转录获得病毒RNA,用脂质体法将病毒RNA转染叙利亚仓鼠肾细胞(BHK-21)包装出病毒粒子,再转接到猴胚胎肾上皮细胞(MACR-145)传代拯救病毒。对拯救的病毒进行RT-PCR扩增、酶切和序列分析鉴定。结果表明,拯救病毒含有不同于亲本病毒的分子标记(病毒基因组14667位人工产生的MluⅠ酶切位点)和插入的标记基因序列。间接免疫荧光试验表明,外源基因NP49在拯救的PRRSV中表达,且能够在PRRSV传代过程中稳定遗传。病毒生长特性比较显示,拯救病毒与亲本病毒在MARC-145细胞上具有相似的增殖特性。本研究构建了含有标记基因PRRSV的感染性克隆并获得了拯救病毒,为进一步开发新型PRRS疫苗提供了有效的反向遗传操作平台。 相似文献
13.
基于模糊系统的开度转速双变量施肥控制序列生成方法 总被引：3，自引：2，他引：1
为协调双变量施肥机车载电脑有限的运算能力与施肥精度和均匀性之间的矛盾,该文提出了一种基于模糊系统的开度转速双变量施肥播种控制序列生成方法。该方法综合考虑前、中、后连续3次施肥播种量的相互影响,实现了实时计算近优的施肥播种控制序列,避免了小施肥量下的大开度小转速现象,减小了外槽轮脉动性的影响。室内试验结果表明,该方法有效地协调了车载电脑运算能力不足和施肥精度等之间的矛盾,平均施肥误差比常用方法减小了4%,同时避免了极端情况下的电机无法驱动排料转轴的情况。田间作业试验证明,使用该方法的施肥机指令响应及时,播撒均匀、准确。 相似文献
14.
Rui Jiang Xiao Li Minghua Zhou Hui Jie Li Ying Zhao Jun Yi 《Soil Science and Plant Nutrition》2016,62(1):1-12
Plastic film mulching has commonly been used for adaptation to water scarcity and for increasing agricultural productivity on the semiarid Loess Plateau of China. However, the effect of plastic film mulching on cropland soil water and thermal regimes on the semiarid Loess Plateau of China is not well understood. This study simultaneously monitored the dynamics of the soil water content and the soil temperature with high resolution in a ridge cultivation system with plastic film mulching (RS) and a flat cultivation system without plastic film mulching (FS) during the maize (Zea mays L.)-growing season. We found that, in general, the soil temperature and soil water content were significantly different among the ridge under RS (RS-ridge), the furrow under RS (RS-furrow) and FS throughout the maize-growing season (P < 0.05). Plastic film mulching increased the near-surface soil temperature by approximately 1°C throughout the study period. RS significantly increased the soil water content during the dry period (May to June), especially within the middle soil layer (30–60 cm), compared to FS. The lowest monthly average soil water content was found at a depth of 30–60 cm layer in FS during the dry period (May and June). The water depletion was found within deeper (100–160 cm) soil layers in May but the water storage in the same layer of FS in June increased although it was the dry period, which differed from RS. The RS practices showed a longer period of water supply from the deeper soil layer (100–160 cm) in May and June for meeting maize water demands during the early growing stage rather than in only May for FS. During June (dry period), the water storage at a depth of 0–60 cm was greater in RS than in FS, and the reverse was true at a depth of 60–160 cm. The results indicate that the dry soil layer at a depth of 30–60 cm formed during June in FS likely reduced water movement from deeper layers to the topsoil layer, and hence constrained the availability of surface soil water for meeting maize water requirements during the early growing stage (dry period). Our study suggests that RS tends to significantly increase surface soil water availability by restraining the formation of a dry soil layer during the early maize-growth stage primarily under dry conditions, and thus enhances maize productivity in the semiarid Loess Plateau of China. 相似文献
15.
Asad Sultan Humera Obaid Sarzamin Khan Inayat ur Rehman Muhammad Kamal Shah Rifat Ullah Khan 《Cereal Chemistry》2015,92(1):50-53
This experiment investigated the effect of a supplementary dietary source (flaxseed) on egg yolk cholesterol contents and fatty acid profile. Flaxseeds at three different levels (5, 10, and 15%) were supplemented to 84 Rhode Island Red × Fayoumi cross layers during 21–24 weeks of age. The birds were divided randomly into four groups. Flaxseeds were supplemented in the diet at the level of 5% (FS‐5), 10% (FS‐10), and 15% (FS‐15), and one group served as the control (FS‐0). Cholesterol content of eggs was not significantly affected by flaxseed. The amount of monounsaturated fatty acids was significantly decreased with the increase in the flaxseed level, and the highest reduction was observed in FS‐15. The highest total polyunsaturated fatty acid (PUFA) and total omega‐3 PUFA were recorded in FS‐15. These findings reflect that unsaturated healthy fatty acid content in poultry eggs can be increased with dietary supplementation of flaxseed. 相似文献
16.
柽柳和梭梭林地土壤呼吸研究 总被引：3，自引：0，他引：3
用便携式气体分析仪（CIRA S-1配备SRC-1呼吸室,PP System s,H itch in,UK）测量了古尔班通古特沙漠南缘阜康荒漠实验站（FS）柽柳丛和北沙窝（ND）梭梭林地冠下和冠间的土壤呼吸及土壤因子。分析表明：冠下和冠间的土壤理化性质存在明显的异质性,冠下土壤呼吸值高于冠间,FS林地的土壤有机质、电导率、全氮、碳酸钙等含量及土壤呼吸值都高于ND林地土壤。土壤呼吸与土壤CaCO3含量有明显的相关性,相关系数为0.89。土壤中CaCO3含量的空间变化对土壤呼吸值的时空变化有一定的影响。掌握土壤因子及土壤呼吸的空间变化,对了解荒漠生态系统过程、土壤微生物与有机质的相互作用及全球气候变化的研究都具有重要的意义。 相似文献
17.
M. T. Ogan 《Water, air, and soil pollution》1994,72(1-4):163-177
The possible advantage of anaerobic membrane filtration (anaerobic-MF) for fecal streptococci (FS) enumeration in tropical (Caribbean) marine recreational waters was evaluated with samples from the beaches of Barbados. Selected samples were also analyzed for fecal coliforms in a comparison. Up to 80% of presumptive FS colonies on m-Enterococcus agar in aerobic-MF were false-positive. Anaerobic-MF reduced presumptive FS and background bacterial counts by 2 to 5 orders of magnitude, increased proportions of FS verification in 54 of the 80 samples, but conferred no obvious advantage in the FC counts. 94 to 100% of blue colonies on MFC agar in 24 hr aerobic- and anaerobic-MF cultures were confirmed as FC. Presumptive and background bacteria in FS counts, proportions of verified FS, and presumptive FC counts correlated with one another. The MF and most probable number (MPN) methods were compared. In a modified MPN confirmation, cultivation of FS colonies in Brain heart infusion agar prior to esculin hydrolysis test gave substantially higher counts than anaerobic-MF at three stations. The overall FS and FC data apparently indicate only low contamination levels occur. Beaches exceeding the region's quality (USEPA) standards are associated with municipal wastewate outfalls. 相似文献
18.
摘要：利用sp18 mAb筛选小鼠睾丸λgt11-cDNA表达文库,命名为sp18（GenBank登录号：AF129872）的阳性克隆由 1 468 bp组成,开放阅读框（open reading frame, ORF）长约429 bp,编码142个氨基酸。起始密码ATG位于第694 bp,终止密码位于第1 122 bp,polyA位于1 245~1 250 bp。核苷酸同源性分析表明,sp18 cDNA的10~651 bp与人视神经Hr44的532~ 1 173 bp、sp18 cDNA的10~816 bp、864~1 444 bp与牛视神经Hr44的1 408~2 208 bp、2 250~2 309 bp的cDNA序列有很高的同源性, 高达98%。推测sp18基因可能是生殖系统存在的一种新基因。sp18编码蛋白的理论分子量约为15.7 kD,有8个N-糖基化位点和12个O-糖基化位点,亲水性分析表明sp18多肽是一种精子跨膜蛋白,跨膜区位于第17~33氨基酸处。将sp18 cDNA亚克隆到原核表达载体pET-30a(+),在E. coli BL21（DE3）中得到诱导表达,其表观分子量约为16.5 kD,表达量约占菌体总蛋白的32%。Western blot分析表明sp18 mAb能识别重组的sp18膜蛋白,表明重组蛋白具有免疫原性,为进一步研究sp18基因的功能打下了基础。 相似文献
19.
Mathematical models employing textural data are commonly used for the estimation of the particle-size distribution (PSD) curve. In this study, two models were used: (1) the model proposed by Fredlund et al. (2000) (F model), and (2) the model proposed by Skaggs et al. (2001) later modified by Fooladmand and Sepaskhah (2006) (FS model). Comparison of the results obtained by these two models, applied for 30 soils from the south of Iran, showed that the F model was better when the model parameters were determined by setting minimum differences between the measured and estimated PSD curve. Also, three equations based on soil textural data were derived for estimating parameters of the F model, and these equations were used to estimate the PSD curve. The results showed that for 10 independent soils from the south of Iran and 50 soils from the UNSODA database, the FS model was better than the F model based on three new derived equations. Therefore, the FS model can be safely used to estimate the PSD curve with different soil textures; however, the F model is appropriate for representation of the measured PSD curve. 相似文献