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21.
以绵羊乳酪蛋白为原料,采用中性蛋白酶水解制备生物活性肽,研究酶解时间对酶解产物水解度和血管紧张素转换酶(angiotensin converting enzyme,ACE)抑制率的影响,利用液相色谱-质谱联用技术对水解产物进行分析和鉴定,筛选具有潜在ACE抑制作用的生物活性肽。结果表明:酶解时间达到300 min时,水解度最高值达9.65%,ACE抑制率为84.55%;当ACE抑制率达到50%时,所需肽段YYQQRP浓度为5~10 μmol/L;利用超高效液相色谱飞行时间质谱仪对酶解后的多肽进行序列分析, 相似文献
22.
试验旨在构建一种更安全有效的新型免疫去势DNA疫苗,通过选取下丘脑-垂体-性腺轴(hypot halamic-pituitary-gonadal axis,HPG)的上游调控基因吻素1(KISS1)和促性腺激素释放激素(GnRH)作为靶标,借助2A肽的自剪切功能,引入平衡致死系统代替抗性基因筛选流程,成功将GnRH和KISS1转入非抗性筛选质粒pVAX-asd中,酶切验证和测序对比验证目的基因的插入方向和序列完全正确。重组质粒转染HeLa细胞,反转录后扩增目的基因结果显示,重组质粒在真核细胞内能够正常转录,保证重组质粒在导入机体后能够正常表达,从而引起特异性免疫反应。将构建成功的质粒转入减毒的猪霍乱沙门氏菌C500中,获得可直接口服免疫的活载体疫苗,酶切和测序结果表明,双表达重组质粒成功导入工程菌中。将活菌疫苗在体外连续传代50次,选取0、2、5、10、20、30、40、50代的菌株进行稳定性研究,生长曲线检测结果表明,工程菌在体外连续传代50次的过程中,其生长特性无明显变化,且与减毒猪霍乱沙门氏菌C500的生长特性一致,未因携带质粒发生明显变化;同时将各代菌液扩增沙门氏菌标志基因(invA)和毒力基因(crp),结果表明多次传代后工程菌仍然具有沙门氏菌的特性,其减毒特性也无变化。各代菌液质粒酶切验证显示,多次传代并不影响质粒的稳定性,重组双表达质粒能够在沙门氏菌C500中维持正常拷贝功能。综上所述,该重组质粒和工程菌疫苗均具有良好的稳定性,可直接应用于动物免疫去势的研究。 相似文献
23.
This experiment aimed to examine the effect of periodical application of bioactive peptides derived from cottonseed (BPC) in comparison with using sub-therapeutic doses of lincomycin and the excessive inclusion of vitamin E on performance, immunity, total antioxidant capacity of serum and intestinal morphology of broiler chickens. A total of 240 one-d-old male broiler chicks with similar initial weight (Ross strain) were randomly assigned to 6 groups (8 chicks/pen): non-treated group (basal diet), basal diet supplemented with 2 mg/kg lincomycin, basal diet supplemented with 50 IU vitamin E, basal diet supplemented with 6 g BPC/kg in starter period, basal diet supplemented with 6 g BPC/kg in starter and grower periods and basal diet supplemented with 6 g BPC/kg throughout the whole experiment. The highest final body weight was obtained in the group supplemented with BPC in starter and grower periods. In the finisher phase, broilers fed the diet containing BPC in the starter period and in the whole trial had significantly (P < 0.05) better feed conversion ratios (FCR). Jejunal villus height was significantly elevated in broilers supplemented with antibiotic (P < 0.001), furthermore it tended to be greater in broilers fed BPC in the starter period. The jejunal villus height-to-crypt depth ratio was significantly (P < 0.01) higher in broilers fed the diet containing antibiotic in comparison to other groups. Humoral immune response against Newcastle disease vaccine tended to be elevated in broilers fed the diet containing BPC in the whole trial (P > 0.05). Broilers supplemented with BPC in starter and grower, and in the whole trial had significantly (P < 0.05) higher antibody titers against sheep red blood cells (SRBC). The highest total antioxidant capacity was obtained in broilers supplemented with the excessive level of vitamin E, furthermore it tended to improve in broilers fed the diet containing BPC in the whole trial. In summary, the results of the study indicated that addition of BPC in broiler diets in the whole trial could improve FCR, immune responses and total antioxidant activity of serum, and BPC could be used in broiler diets as an alternative to in-feed antibiotics. 相似文献
24.
Hiroshi BANNAI Manabu NEMOTO Koji TSUJIMURA Takashi YAMANAKA Ken MAEDA Takashi KONDO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(2):309-311
To increase the sensitivity of an enzyme-linked immunosorbent assay (ELISA) for equine herpesvirus type 4
(EHV-4) that uses a 12-mer peptide of glycoprotein G (gG4-12-mer: MKNNPIYSEGSL) [4], we used a longer peptide consisting of a 24-mer repeat sequence (gG4-24-mer:
MKNNPIYSEGSLMLNVQHDDSIHT) as an antigen. Sera of horses experimentally infected with EHV-4 reacted much more
strongly to the gG4-24-mer peptide than to the gG4-12-mer peptide. We used peptide ELISAs to test paired sera
from horses naturally infected with EHV-4 (n=40). gG4-24-mer ELISA detected 37 positive samples (92.5%),
whereas gG4-12-mer ELISA detected only 28 (70.0%). gG4-24-mer ELISA was much more sensitive than gG4-12-mer
ELISA. 相似文献
25.
饲粮能量和蛋白质水平对滩羊小肠中小肽和氨基酸转运载体mRNA表达量的影响 总被引:2,自引:0,他引:2
本试验旨在研究饲粮能量和蛋白质水平对滩羊小肠中小肽和氨基酸转运载体mRNA表达量的影响。选取112只健康、体重相近的滩羊,随机分成4组,每组4个重复,每个重复7只羊。标准水平的饲粮能量和蛋白质水平参考《肉羊饲养标准》(NY/T 816—2004),各组试验滩羊分别饲喂不同能量和蛋白质水平饲粮:0.84×标准水平(Ⅰ组)、0.96×标准水平(Ⅱ组)、1.08×标准水平(Ⅲ组)和1.20×标准水平(Ⅳ组)。试验根据羊体重分2个阶段:29~35 kg和36~40 kg。于每个阶段末,每个重复屠宰1只试验羊,取其小肠组织样,运用实时荧光定量PCR技术,研究小肽转运载体1(Pep T1)、y+型氨基酸转运载体1(CAT1)、兴奋性氨基酸转运载体3(EAAT3)mRNA表达量的变化。结果表明:1)在29~35 kg阶段末,小肠中Pep T1 mRNA的表达量随着饲粮能量和蛋白质水平的提高呈先下降再上升的趋势,Ⅱ组显著低于其他3组(P0.05);Ⅳ组小肠中CAT1 mRNA的表达量显著高于其他3组(P0.05);Ⅲ组小肠中EAAT3mRNA的表达量显著高于其他3组(P0.05)。2)在36~40 kg阶段末,Ⅱ组小肠中Pep T1mRNA的表达量显著高于其他3组(P0.05);Ⅱ组小肠中CAT1 mRNA的表达量显著高于Ⅲ组(P0.05);小肠中EAAT3 mRNA的表达量随着饲粮能量和蛋白质水平的提高呈上升趋势,Ⅲ组和Ⅳ组小肠中EAAT3 mRNA的表达量显著高于Ⅰ组和Ⅱ组(P0.05)。由此可见,饲粮能量和蛋白质水平会影响滩羊小肠中Pep T1、CAT1、EAAT3 mRNA的表达量,使机体对小肽和氨基酸的吸收利用率随之改变,以适应滩羊的生长发育。 相似文献
26.
27.
噬菌体展示三肽囊素模拟肽的研究 总被引:4,自引:0,他引:4
本研究分离纯化抗Bursin单克隆抗体(2F9-4/HU2),利用噬菌体环7肽库筛选抗Bursin单克隆抗体(2F9-4/HU2)结合肽,测定阳性克隆ssDNA序列,并与Bursin序列进行同源性分析,通过ELISA法检测其结合活性和竞争ELISA法检测其抗原性,以Busin为对照,验证阳性克隆(№4)在鸡体内的生物学效应.序列分析表明,"LNXT"短肽序列可能为结合抗Bursin单克隆抗体(2F9-4/HU2)的保守序列,但与Bursin无同源性;噬菌体(№1)在序列中含有K、H、G.ELISA和竞争ELISA检测,表明Bursin对阳性克隆1、4、5、6号和抗Bursin单克隆抗体(2F9-4/HU2)的结合有一定的抑制作用:生物学活性初步验证表明,阳性克隆(№4)与Bursin具有类似作用,能促进抗体产生,有望成为一种模拟Bursin新型的免疫增强剂. 相似文献
28.
Hiromi Takahashi Chieko Kai Yasuhiro Yoshikawa Kazuya Yamanouchi 《Comparative immunology, microbiology and infectious diseases》1995,18(4):275-281
Pseudorabies virus (PRV) was inoculated intraocularly into BALB/c mice, and the distribution pattern of cells positive for several neurotransmitters and viral antigens in the eyeball, trigeminal nerve ganglia, and superior cervical ganglia was examined immunohistochemically to clarify the neural route of the virus spread. In the eyeball, substance P (SP)- and calcitonin gene-related peptide (CGRP)-positive cells were detected in the ipsilateral iris and ciliary body, neuropeptide tyrosine (NPY)-positive cells were detected in the choloid membrane, and tyrosine hydroxylase (TH)-positive cells were detected in the ipsilateral inner nuclear layer of the retina; all these cells contained viral antigens. In the superior cervical ganglia, viral antigen-positive cells containing TH or NPY were found at bilateral sites. In the trigeminal nerve ganglia, viral antigen-positive cells containing SP or CGRP were found at bilateral sites. These findings indicated that the SP- and CGRP-positive ganglion cells of the trigeminal nerve ganglia innervating the iris or ciliary body, and the NPY-positive ganglion cells of the superior cervical ganglia innervating the iris, ciliary body, and choroid membrane served as the route for the virus spread. These findings also suggested that dopaminergic neurons, such as the TH-positive retinal cells and TH-positive ganglion cells of the superior cervical ganglia, served as the route for virus spread. 相似文献
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