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1.
为进一步研究家蚕胰凝乳蛋白酶抑制剂基因SCI-SB的功能和作用机理,利用RT-PCR技术,从家蚕总RNA中扩增到家蚕SCI-SB基因片段,构建了含有GST标签的融合表达质粒pGEX4T-1-SCI-SB。采用pGEX融合蛋白表达系统,在大肠杆菌BL21中得到以包涵体形式存在的重组GST融合蛋白质,表达量可占菌体蛋白的15%。将包涵体变性、复性处理以后,进一步利用GSTTrapFF亲合层析一步获得了重组蛋白。将该重组蛋白免疫新西兰纯种大白兔,获得效价高的多克隆抗体。胰凝乳蛋白酶活性抑制实验结果表明该重组蛋白具有一定的酶活抑制作用。 相似文献
2.
In vitro experiments were carried out to test the efficacy of plant activator Acibenzolar-S-methyl (ASM, a benzothiadiazole derivative; trade name Bion 50WG) against rhizome rot disease of turmeric (Curcuma longa L.) caused by Pythium aphanidermatum. The plant activator was applied as a liquid rhizome pre-treatment followed by inoculation with P. aphanidermatum. Cell death, activities of pathogenesis related (PR) proteins such as cysteine protease (EC 3.4.22), peroxidases (EC 1.11.1.7) both soluble and ionically bound (IB), trypsin inhibitor (EC 3.4.21.1) and chymotrypsin inhibitor (EC 3.4.21.4) were monitored. Rhizome pre-treatment was effective in controlling P. aphanidermatum infection. Anatomical observation of turmeric rhizomes indicated the presence of calcium oxalate deposits in infected tissue and an accumulation of starch grains in response to infection by P. aphanidermatum. Pathogen infection also induced new basic polypeptides corresponding to 18.0 and 41.0 kDa. Induction of protease, protease inhibitors, soluble and ionically bound peroxidase activity was observed after ASM pre-treatment and P. aphanidermatum infection. ASM treatment also enhanced activities of proteases and peroxidase in rhizomes already infected with P. aphanidermatum. Increases in enzyme activities and protease inhibitors occurred much more rapidly and were enhanced in P. aphanidermatum infected rhizomes that were previously treated with ASM suggesting that increased activities of peroxidases and protease inhibitors may play a key role in restricting the development of disease symptoms on the rhizomes infected with P. aphanidermatum as evidenced by a reduction in cell death. Hence, pretreatment with ASM suppress the P. aphanidermatum induced oxidative damage through higher accumulation of peroxidases and induced defense through activities of protease inhibitors thereby, protected turmeric rhizomes from rhizome rot disease. 相似文献
3.
将12窝杜洛克×长白×约克夏三元杂交新生仔猪随机分为4组,分别于17、21、28和35日龄断奶后,测定断奶前后胰腺组织和十二指肠、空肠、回肠的食糜及粘膜中胰糜蛋白酶的相对活性、比活和总活性。结果为:哺乳仔猪在断奶前胰腺糜蛋白酶活性保持较高水平,22日龄达到最高值,29日龄时明显下降（P<0.05）。而仔猪断奶后12h各组胰腺糜蛋白酶的相对活性和比活均显著提高,较同期哺乳仔猪高,随后降低,且在断奶后2～3周内均保持较低水平。18至50日龄间,哺乳仔猪及断奶仔猪胰腺中糜蛋白酶总活性在哺乳期和断奶后并没有随胰腺重量的增加而增加（P<0.05）。哺乳期空肠中糜蛋白酶活性的变化模式与胰腺的大体一致。若断奶日龄早于28日龄时,断奶后1周空肠中糜蛋白酶的活性降低。而28、35日龄断奶,断奶后这两种酶的活性不受影响,而且断奶日龄越早,降低后恢复至正常值的时间越长。十二指肠糜蛋白酶活性在断奶后的变化情况基本与空肠相似,但是回肠中却没能有规律地反映糜蛋白酶活性与日龄之间的关系。 相似文献
4.
V. R. Devaraj N. H. Manjunath 《Plant foods for human nutrition (Dordrecht, Netherlands)》1995,48(2):107-112
Proteinase inhibitory activity in ten different varieties ofDolichos lablab perpureus. L. was determined. All the varieties tested exhibited appreciable level of proteinase inhibitory activity (PIA). The trypsin inhibitory activity (TIA) (Mean:20170 TIU/g) was relatively higher than the chymotrypsin inhibitory activity (CIA) (Mean: 15380 CIU/g). Effect of temperature and cooking on PIA was studied. The nature of cooking medium and duration of cooking had profound effect on the PIA. The dry fried seeds lost their PIA very rapidly (91% in 20 min). Seeds cooked in slightly alkaline medium lost their PIA quickly (89% in 30 min) compared to those cooked in acidic (80% in 30 min) and neutral pH (83% in 30 min). The PIA in green pods was also determined and they had only one third of the PIA (8200 TIU/g and 8125 CIU/g) found in the dry seeds. 相似文献
5.
V. H. Mulimani S. Vadiraj 《Plant foods for human nutrition (Dordrecht, Netherlands)》1994,46(1):27-31
Changes in trypsin and chymotrypsin inhibitory activity of sorghum (Sorghum bicolor L. Moench) seeds on soaking in distilled water, different salt solutions and mixed salt solution were investigated. A greater reduction in proteinase inhibitory activity was observed on soaking in salt solution than on soaking in distilled water. Maximum loss of trypsin and chymotrypsin inhibitory activity was observed on soaking seeds in mixed salt solution. 相似文献
6.
0～3周肉仔鸡消化道酶发育规律的研究 总被引：25，自引：2，他引：25
本试验以０～３周Ａｖａｉｎ商品代肉仔鸡公雏为研究对象,测试０,４,７,１０,１４,１７和２１日龄肉仔鸡体重,消化道器官重量,胰腺和肠道食糜中淀粉酶、胰蛋白酶、糜蛋白酶和脂肪酶活性的变化,基本摸清了０～３周肉仔鸡体重、消化器官的生长规律及消化道酶的发育规律。试验结果表明：（１）肉仔鸡在４～１０日龄日均相对生长率达到最大值（平均每天增重达１９％）,１０日龄后下降,到１４～２１日龄,基本维持在１３％～１４％左右;（２）各器官的相对重量（ｇ／ｋｇ体重）达到峰值的时间并不一致,腺胃和肌胃的相对重量在４～７天为最高,肝和胰在７～１０天,小肠在７天左右;（３）从异速生长的指标看,肌胃的最大增长倍数是体重增长倍数的１．５倍,腺胃是２倍,肝脏是２．３倍,小肠是３倍,胰腺达６倍;（４）小肠内容物的相对重量（ｇ／ｋｇ体重）在４～１０日龄达到最大值,以后呈下降趋势;（５）胰腺和肠道的消化酶活性（活性单位／ｋｇ体重）基本上随着日龄的升高而上升。在胰腺,淀粉酶、胰蛋白酶、糜蛋白酶和脂肪酶活性均在１０,７,２１和１０日龄达到峰值。在肠道,淀粉酶、胰蛋白酶、糜蛋白酶和脂肪酶活性均在１０日龄达到峰值。从消化道酶的分泌情况看,肉仔鸡早期消化酶 相似文献
7.
AIM: To investigate the diverse pathways of local angiotensin Ⅱ (Ang Ⅱ) generation in islets of Langerhans in Syrian golden hamsters with dyslipidemia. METHODS: The Syrian golden hamsters were fed with high-fat diet to induce dyslipidemia. Purified islet cells from dyslipidemia and normal Syrian hamsters were prepared and divided into control group, captopril group, chymotrypsin endostatin group, aprotinin group, α-antitrypsin group and captopril+chymotrypsin endostatin group by adding respective reagents into the cultured cells after treated with angiotensin I. The Ang Ⅱ levels in the supernatants of each group were examined by ELISA.RESULTS: Compared with the control animals, Ang Ⅱ levels decreased in all groups with interventions. Compared with the normal hamsters islet cells, the Ang Ⅱ levels in captopril group, chymotrypsin endostatin group, α-antitrypsin group, aprotinin group and captopril+chymotrypsin endostatin group were decreased by 39.98%, 50.10% (P<0.01), 23.04%, 20.85% (P<0.05) and 82.78% (P<0.01), respectively. Compared with high-fat group, the corresponding data were 42.12%, 56.96% (P<0.01), 26.11%, 22.68% (P<0.05) and 83.59% (P<0.01), respectively. The levels of Ang Ⅱ in chymotrypsin group between normal and high-fat diet hamsters were significantly different (P<0.05). CONCLUSION: Under the condition of dyslipidemia, the classic angiotensin-converting enzyme-based pathway and chymotrypsin pathway are still the main approaches of producing Ang Ⅱ in male Syrian hamster islet to produce angiotensin. The effect of chymotrypsin endostatin is comparatively stronger in inhibiting the production of local Ang Ⅱ than the effect of angiotensin-converting enzyme inhibitor. 相似文献
8.
ZHANG Hong-fu LI Chang-zhong GU Xian-hong FANG Lu MA Yong-xi GONG Li-ming Feng Guang-ming 《中国农业科学(英文版)》2002,1(3):338-341
Twelve litters of new born piglets were divided randomly into groups Ⅰ, Ⅱ, Ⅲ and Ⅳ, and weaned at 17, 21, 28 and 35 days of age, respectively, to determine the effect of weaning age on pancreatic and intestinal chymotrypsin activity in piglets. The results showed that the relative and specific activity of pancreatic chymotrypsin increased significantly 12h after weaning compared with that of sucking piglets at the same age and then decreased and remained low for 2 - 3 weeks after weaning. Total pancreatic chymotrypsin activity had no change with the increase of pancreas weight during day 18 - 50 regardless of weaning (P ＞0.05). Chymotrypsin activity of jejunum had the same change as that of pancreas during sulking stage.Jejunum chymotrypin activity decreased in the first week post-weaning in the piglets weaned before day 28, but had no change in groups weaned at 28 or 35 days. The earlier the weaning age, the longer the restoring time.Chymotrypsin activity in jejunum is more sensitive to the effects of weaning age than in duodenum and ileum. 相似文献
9.
Proteinase inhibitors (AsPIs) with high activity against serine proteinases were purified from seeds of the tree legume, Acacia senegal by ammonium sulfate precipitation followed by DEAE-Sephadex A-25 column and evaluated against Helicoverpa armigera larvae by in vitro and in vivo methods. The molecular weight of AsPIs was found to be approximately 19.58 ± 1.00 and 21.23 ± 1.00 kDa for PI and 18.16 ± 1.00 kDa for PII on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The AsPIs (5 μg/ml) inhibited approximately 70% of midgut trypsin and 61% of elastase-like chymotrypsin. In vitro studies showed that AsPIs have remarkable inhibitory activity towards total gut proteolytic enzymes followed by trypsin and chymotrypsin. The IC50 of AsPIs for midgut trypsin was 0.1 μg/ml and for chymotrypsin was 2.0 μg/ml. The inhibition of gut proteinase enzymes was of the non-competitive type. In larval feeding studies, AsPIs were found to retard growth and development of H. armigera and also affects the fecundity of the pest. The results advocate the use of AsPIs in transgenic technology to develop plant resistance to H. armigera. 相似文献
10.
用PCR方法从家蚕基因组DNA扩增了家蚕胰凝乳蛋白酶抑制剂b1基因(CIb1)4个不同长度的启动子片段,构建由其驱动的萤火虫荧光素酶基因报告质粒,在脂质体介导下转染家蚕BmN细胞,体外分析了该基因启动子的活性。结果表明,家蚕CIb1基因启动子在BmN细胞中有微弱的转录活性;野生型BmNPV感染能增强启动子活性;hr3增强的不同长度CIb1基因启动子片段的活性有显著差异,提示转录起始位点上游-74～-1nt包含了启动子的基本元件,而在-687～-465nt、-465～-317nt和-317～-74nt存在着主要的顺式元件。试验结果有助于阐明CIb1的表达调控机理和对家蚕天然性免疫的理解。 相似文献