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1.
绵羊甘露聚糖结合凝集素（MBL）基因的克隆及序列分析 总被引：2，自引：1，他引：1
甘露聚糖结合凝集素(MBL),又称甘露聚糖结合蛋白(mannan/mannose-binding protein,MBP),是C型凝集素超级家族中胶凝素家族成员.目前,关于MBL的研究在人、小鼠和大鼠上已经相当详尽,在牛、猪、马、弥猴、黑猩猩、狒狒、兔和鸡上也有一定的研究,除鸡外,其余的都已发现2种MBL:MBL-A(血清型)和MBL-C(肝型),但在绵羊上仍为空白.本实验以绵羊的基因组DNA为模板,借助人、牛MBL的保守序列来获得绵羊MBL基因片段. 相似文献
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利用抑制消减杂交技术研究同一只西农萨能奶山羊泌乳中期和末期的乳腺组织差异表达基因,以泌乳中期乳腺cD-NA为测试组,末期乳腺cDNA为驱动组,构建消减文库,得到山羊(Capra hircus)嗜乳脂蛋白基因的部分序列。根据牛(Bos)和绵羊(Ovisaries L.)基因组序列进行电子拼接,并设计引物,得到山羊嗜乳脂蛋白基因的CDs区全序列,应用RT-PCR技术从山羊乳腺组织总RNA中扩增克隆了山羊嗜乳脂蛋白基因CDs区,命名为gBTN1A1,并登录GenBank(EF102891)。gBTN1A1基因开放读码框由1581个碱基组成,编码526个氨基酸,前26个氨基酸为推定的信号肽区域。gBTN1A1基因核苷酸序列与牛(NM-174508)、人(NM-001732)和鼠(AK145168)的同源性分别为97%、88%和84%,氨基酸序列的同源性分别为96%、84%和70%。其二级结构、跨膜区域及信号肽分析均与牛、人和鼠的BTN1A1基因相似。 相似文献
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山羊FSHR基因第10外显子的PCR-SSCP检测及其序列分析 总被引：4，自引：0，他引：4
利用PCR-SSCP技术检测山羊(Caprahircus)包括西农萨能奶山羊、关中奶山羊、陕南白山羊、安哥拉山羊和波尔山羊173个个体FSHR基因第10外显子的单核苷酸多态性(SNP)。结果未发现SNP位点。测序后获得山羊FSHR基因第10外显子的核苷酸序列,并在NCBI数据库中获得GenBank登录No.DQ069909和DQ069910。通过DNA序列分析发现,FSHR基因第10外显子第120位碱基不存在C→T的转换,也不存在颠换等其它遗传变化。山羊、绵羊(OvisariesL.)和普通牛(Bostaurus)FSHR基因第10外显子序列同源性比较和聚类分析结果表明,山羊、普通牛和绵羊该部分序列的相似性最高为99.3%;在物种间比较中,绵羊和普通牛纯合子该基因外显子序列的不相似性最高为3.4%;据FSHR基因外显子序列构建的分子系统树结果显示,山羊、绵羊和普通牛物种内的个体各自聚为一类;山羊和绵羊先聚为一类,然后再与普通牛聚为一类。提示FSHR基因第10外显子的核苷酸序列适合于物种间的动物分子树的构建。 相似文献
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蓝贤勇；陈宏；潘传英；雷初朝；张永德；于姣 《农业生物技术学报》2006,14(4):484-488
利用PCR-SSCP技术检测山羊(Capra hircus)包括西农萨能奶山羊、关中奶山羊、陕南白山羊、安哥拉山羊和波尔山羊173个个体FSHR基因第10外显子的单核苷酸多态性 (SNP)。结果未发现SNP位点。测序后获得山羊FSHR基因第10外显子的核苷酸序列， 并在NCBI数据库中获得GenBank登录No.DQ069909和DQ069910。通过DNA序列分析发现， FSHR基因第10外显子第120位碱基不存在C→T的转换，也不存在颠换等其它遗传变化。山羊、绵羊 (Ovisaries L.) 和普通牛( Bos taurus) FSHR基因第10外显子序列同源性比较和聚类分析结果表明，山羊、普通牛和绵羊该部分序列的相似性最高为99.3%；在物种间比较中，绵羊和普通牛纯合子该基因外显子序列的不相似性最高为3.4%；据FSHR基因外显子序列构建的分子系统树结果显示，山羊、绵羊和普通牛物种内的个体各自聚为一类；山羊和绵羊先聚为一类，然后再与普通牛聚为一类。提示FSHR基因第10外显子的核苷酸序列适合于物种间的动物分子树的构建。 相似文献
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通过PCR扩增,从融安金柑基因组DNA中克隆出一条2 361 bp的DNA片段,该DNA克隆含有1个2 081 bp的LEAFY同源基因全长序列(FcLFY)。金柑LEAFY同源基因包含2个内含子和3个外显子,编码398个氨基酸。比对结果表明,金柑LEAFY同源基因与甜橙、枳的LEAFY同源基因的核苷酸和氨基酸序列的同源性均为98%。该研究为今后从分子水平上研究金柑开花调控机理打下了坚实的基础。 相似文献
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猪骨形成蛋白15基因编码区序列的克隆及测序研究 总被引：3，自引：0，他引：3
基于比较基因组学方法,选择大白猪和梅山猪作为试验材料,根据人、小鼠和猪的BM P 15基因设计并合成4对引物,进行基因组DNA的PCR扩增、克隆、测序,用BLA ST软件进行DNA序列排列,获得包含猪BM P 15基因外显子1(exon1)和外显子2(exon2)的全部编码区序列。用Pa irw ise BLA ST软件,将大白猪、梅山猪BM P 1 5基因编码区序列进行比较,在外显子2区域发现了一个SNP位点,位于编码区第390个核苷酸处,大白猪为T,梅山猪为A,且该位点导致了限制性内切酶Sp eⅠ酶切位点发生了改变。建立了猪BM P 15基因基于内切酶Sp eⅠ的PCR-RFLP多态性检测技术,发现猪BM P 15基因有3种基因型(BM P 15AA、BM P 15AB、BM P 15BB)。 相似文献
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鹅脂联素基因的克隆、序列分析及组织表达 总被引：5，自引：1，他引：4
根据GenBank发表的鸡和鸭等物种的脂联素基因序列的同源保守区域,设计了3对特异性引物,以鹅(Anser domeatica)血液基因组DNA为模板,经扩增、测序及拼接得到了部分DNA序列,全长1062bp,包含2个外显子和1个内含子(GenBank Accession No.EU370686)。该基因编码区长738bp,编码245个氨基酸,编码区核苷酸序列及所编码氨基酸序列与鸭的同源性最高,分别为94.85%和95.51%,与鸡的同源性次之,而与哺乳类的同源性较低。蛋白预测的分子量和等电点分别为26603.3D和5.19,与鸡、鸭、人及小鼠的脂联素蛋白相似。半定量PCR结果显示鹅脂联素基因在骨骼肌、脂肪、心脏和肌胃中高度表达,在小肠、腺胃、肾和肺中中度表达,而在肝脏、脾、卵巢和间脑中低度表达。 相似文献
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张丽娟;罗军;武会娟;刘俊霞;韩雪峰;张宁;王海滨;单翠燕;余刚 《农业生物技术学报》2007,15(5)
利用抑制消减杂交技术研究西农萨能羊泌乳中期和末期的乳腺组织差异表达基因，构建消减文库，得到山羊嗜乳脂蛋白的部分序列，根据牛和绵羊基因组序列进行电子拼接，设计引物，得到山羊嗜乳脂蛋白的CDs区全序列，应用RT-PCR技术从山羊乳腺组织总RNA中扩增克隆了山羊嗜乳脂蛋白基因CDs区，命名为gBTN1A1,并登录Genbank（EF102891）。gBTN1A1全基因由7个外显子和6个内含子组成，开放读码框由1581个碱基,编码526个氨基酸，gBTN1A1基因核苷酸序列与牛、人和鼠的同源性为97%, 88%, 84%，蛋白质序列的同源性为96%, 84% and 70%。其二级结构、跨膜区域及信号肽分析与牛、人和鼠相似，所以推测gBTN1A1与乳脂肪球的分泌密切相关，根据其在泌乳期表达丰度的差异推测其可能影响山羊产奶量。 相似文献
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以"三月红"荔枝基因组DNA为模板,用Cu*ZnSOD基因特异寡聚核苷酸为引物,进行PCR扩增,得到特异基因片段,将其克隆到pGEMT载体上,转化感受态大肠杆菌TG1中,对转化子中重组pGEMT上的Cu*ZnSOD基因片段的PCR扩增检测和序列分析,表明克隆成功;该基因片段有478个核苷酸,由4个外显子和3个内含子组成;外显子由234个核苷酸组成,编码78个氨基酸;该基因片段编码的氨基酸序列与水稻、玉米、番茄、大白菜和松树的Cu*ZnSOD基因编码的氨基酸序列的同源性分别为79.5%、73.1%、73.1%、71.8%和75.7%. 相似文献
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绵羊C3d基因克隆及分子特征 总被引：1，自引：0，他引：1
本实验首次从绵羊肝脏组织克隆到了补体C3d的基因并对其核苷酸序列和推导的氨基酸序列以及蛋白结构进行了分析。绵羊补体C3d基因的编码区含有909个核苷酸,编码303个氨基酸残基,绵羊与人、牛、山羊、野猪、金仓鼠、小鼠、褐鼠、大袋鼠、兔和原鸡推导出的氨基酸序列的一致性分别为80.4% ,94.7% ,96.9% ,82.8% ,81.1% ,80.6% ,80.2%,71.0% ,75.2%和60.3%。高等动物中,绵羊与原鸡的一致性最低为60.3%,而与其他动物之间的一致性则较高为71.0%-96.9%。通过生物学软件分析发现绵羊C3d蛋白有2个蛋白激酶C磷酸化位点,1个酪蛋白激酶Ⅱ磷酸化位点,1个酪氨酸激酶磷酸化位点,1个十四(烷)酰化位点。绵羊C3d二级结构中螺旋和转角交替出现,其中α-螺旋占55.12%、β-折叠为2.31%,转角区域为42.57%,这样的结构有利于其桶状结构的形成。通过ESyPred3D同源建模预测可知绵羊C3d三级结构与人C3d一样也形成由核心和外层构成的桶状分子结构。本实验不仅为C3d在家畜上的免疫学基础研究提供实验依据,而且为构建高免疫原性的基因疫苗新技术奠定基础。 相似文献
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Identification of oxidation products of (-)-epigallocatechin gallate and (-)-epigallocatechin with H(2)O(2) 总被引：4，自引：0，他引：4
Zhu N Huang TC Yu Y LaVoie EJ Yang CS Ho CT 《Journal of agricultural and food chemistry》2000,48(4):979-981
(-)-Epigallocatechin gallate (EGCG) and (-)-epigallocatechin (EGC) are two important antioxidants in tea. They also display some antitumor activities, and these activities are believed to be mainly due to their antioxidative effects. However, the specific mechanisms of antioxidant action of tea catechins remain unclear. In this study are isolated and identified two novel reaction products of EGCG and one product of EGC when they were reacted separately with H(2)O(2). These products are formed by the oxidation and decarboxylation of the A ring in the catechin molecule. This study provides unequivocal proof that the A ring of EGCG and EGC may also be an antioxidant site. This study also indicates an additional reaction pathway for the oxidation chemistry of tea catechins. 相似文献
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W R Dantzer J Hopper K Mullin S Hendrich P A Murphy 《Journal of agricultural and food chemistry》1999,47(10):4291-4296
14C-Fumonisin B(1) (FB(1)) was produced by Fusarium proliferatum M-5991 in modified Myro liquid medium and purified to >95% purity with a specific activity of 1.7 mCi/mmol. Nine male and nine female F344/N rats were each dosed by gavage with 0.69 micromol of (14)C-FB(1), (14)C-hydrolyzed FB(1), or (14)C-FB(1)-fructose/kg body weight. Urinary excretion of (14)C-FB(1) and (14)C-FB(1)-fructose was 0.5% and 4.4% of the total dose, respectively, and was similar between male and female rats. Urinary excretion of (14)C-hydrolyzed HFB(1) was significantly greater (P > 0.05) in female rats as compared with male rats (17.3% vs 12.8% of the total dose, respectively). There were no significant (P > 0.05) differences in biliary excretion of the three fumonisin compounds with a mean of 1. 4% of the dose excreted at 4 h after dosing. Lesser amounts continued to be excreted up to 9.25 h after dosing. Although biliary excretion of the (14)C-FB(1), (14)C-hydrolyzed FB(1), and (14)C-FB(1)-fructose was similar, increased urinary excretion of the (14)C-hydrolyzed FB(1) as compared to (14)C-FB(1) and (14)C-FB(1)-fructose indicated a greater absorption of the hydrolyzed form. 相似文献
13.
Roy A 《Journal of agricultural and food chemistry》1999,47(12):5209-5210
Terpinolene oxide, a monoterpene belonging to the p-menthane group, is easily derived from naturally abundant (R)-limonene. It was isomerized with montmorillonite clay catalyst to karahanaenone (2,2, 5-trimethylcyclohept-4-en-1-one) by ring enlargement. The enantiomers of the corresponding alcohol, karahanaenol (2,2, 5-trimethylcyclohept-4-en-1- ol), known for their individual organoleptic properties, were resolved through Pseudomonas cepacia lipase mediated enantiospecific alcoholysis of its acetate derivative. 相似文献
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The presence of ethylenediamine-N-(o-hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA) as the second largest component in commercial EDDHA iron chelates has recently been demonstrated. Here is reported the speciation of o,p-EDDHA by the application of a novel methodology through the determination of the complexing capacity, protonation, and Ca(2+), Mg(2+), Cu(2+), and Fe(3+) stability constants. The pM values and species distribution in solution, hydroponic, and soil conditions were obtained. Due to the para position of one phenol group in o,p-EDDHA, the protonation constants and Ca and Mg stability constants have different values from those of o,o-EDDHA and p,p-EDDHA regioisomers. o,p-EDDHA/Fe(3+) stability constants are higher than those of EDTA/Fe(3+) but lower than those of o,o-EDDHA/Fe(3+). The sequence obtained for pFe is o,o-EDDHA/Fe(3+) >/= o,p-EDDHA/Fe(3+) > EDTA/Fe(3+). o,p-EDDHA/Fe(3+) can be used as an iron chelate in hydroponic conditions. Also, it can be used in soils with limited Cu availability. 相似文献
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S. R. Verma J. P. S. Arora J. S. Shankar R. Chand 《Water, air, and soil pollution》1987,36(3-4):247-257
The effect of protein oxovanadium(V) ion concentration and pH on the ratio of diffusion current (id/id0) was studied in vanadium(V) ovalbumin-S and denatured ovalbumin systems. In both the cases marked decrease in diffusion current was observed at the respective pH values, indicating that binding takes place with cationic groups of the proteins. The binding sites (n) were found to be pH dependent. The uniformity of logK and ΔG 0 value at all pH values indicated the involvement of same sites in interaction. Furthermore, the linear scatchard plots in both the systems supported the involvement of single class of independent sites in oxovanadium(V) anion interaction. The difference in binding sites (n) has been attributed to the folded structure of ovalbumin-S while unfolded one of denatured ovalbumin. 相似文献
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The efficiency of As(III) oxidation by MnO2, and retention of oxidation products varies with system pH. Maximum retention by hydrous Mn(IV) oxide occurs at pH < 5, declining at higher pH to about half total As at pH 10. The adsorption capacities of pyrolusite and cryptomelane at pH ～6.5 for As(V) species were 10 and 25 mmol kg?1, respectively. HMO surface saturation (～10 mmol kg?1) was reached with equilibrium As(V) levels of 5 to 8 × 10?6 M but this was supplemented at higher levels by an absorption process where uptake increased linearly with concentration (e.g., 68 mmol kg?1 with 2 × 10?5 M As(V)). Added As(III) was avidly oxidized and most product retained at pH 3. At higher pH increasing amounts of As(III) remained unoxidized due to initial reactions apparently blocking access to internal pores. Added Na+ reduced the amount of As retained by the HMO, with the phosphate salt having a significant effect. Extraction studies confirmed that most As could be released by exposure to reducing agents or chelating agents (EDTA). The environmental significance of the results has been considered. 相似文献