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The apoB gene polymorphism on Xba I site of 53 6 unrelated Chinese Han people,aged 25~64 years,was analyzed using PCR and Re striction Fragment Length Polymorphismmethod;serum apoA I,apoB,total chole s terol,triglyceride,HDL cholesterolwere measured,and LDL choleste roland non-HDL cholesterolwere calculated.

采用聚 合酶链反应结合限制性片段长度多态性分析的方法,分析536名25~64岁的无血缘关系的汉族人的apoB基因Xba I位点多态性,并测定其血清apoA I、apoB、总胆固醇、甘油三酯、高密度脂蛋白胆固醇,并计算低密度脂蛋白胆固醇LDL-C)及非高密度脂蛋白胆固醇。

METHODS: MyoD cDNA fragments were extracted from plasmids pEMSV-MyoD with polymerase chain reaction, and PCR was used to clone the whole-length gene of MyoD. After adding CACC sequence at 5' end, MyoD gene was cloned by orient topology into transfer ventor, pENTR/D-TOPO. Objective gene was transferred into adenoviral expression vector DNA via pENTR/D-TOPO vector. The recombinant adenoviral vectors transfected into HEK293A cells by using lipofectamine were packaged and amplified.

从pEMSV-MyoD质粒上用聚合酶链反应法扩增出MyoD cDNA片段,再通过聚合酶链反应使MyoD基因加上CACC序列接头,经过定向拓扑克隆使目的基因连接到转移载体上,再通过LR酶促反应,将目的基因转移到腺病毒表达载体DNA上,获得MyoD基因重组的腺病毒DNA,用脂质体转染法转染HEK293A细胞,包装扩增出MyoD基因重组的腺病毒。

Congenital ichthyosis and psoriasis. Therefore, further investigation into the relationship between 12R-LOX, eLOX3 and skin barrier, as well as the development of novel drugs targeting on these genes would provide new ideas and methods for the treatment of xerosis and development of moisturizers.

深入研究12R-脂氧合酶和表皮型脂氧合酶3与皮肤屏障的关系,以及研发选择性作用于二者基因靶点的新型药物将为某些干燥性皮肤病的治疗和保湿类化妆品的开发提供新的思路和方法。

We detected fasting plasma glucose by glucose oxidase, fasting insulin by chemiluminescent immunoassay, triglyceride and high density lipoprotein-cholesterol by zymology, tumor necrosis factor-αby enzyme linked immunosorbent assay, the reverse of the product of FPG and FIns is used as the insulin sensitivity index, that is, ISI=1/FPG×Fins.

血糖测定采用葡萄糖氧化酶法;胰岛素测定采用化学发光法;血脂(甘油三酯TG、高密度脂蛋白HDL-C)测定采用酶法;肿瘤坏死因子-α测定采用双抗体夹心酶联免疫法;胰岛素敏感指数采用空腹血糖与空腹胰岛素乘积的倒数,即ISI=1/FPG ×FIns。

The substrate specificity test showed that the lipolytic enzyme from strain DF-B6 was an esterase, and not a lipase.

底物特异性实验确定DF-B6菌株分泌的脂裂解酶为酯酶,而非脂肪酶。

The results showed that ACP positive reactions were mainly deposited in lysosome, nucellar cell, cellular endomembrane, mitochondriurn, and endoplasmic reticulum in four tissues of the healthy shrimp. Furthermore, ACP activity was located in microvilli, and around partly lipid droplet in liver. AKP positive reactions were mainly deposited in cell membrane, nucellar cell, lysosome. endoplasmic reticulum and cellular endomembrane in four tissues.

结果显示,在健康的对虾体内,ACP依次出现在各组织中的溶酶体、细胞核、细胞内膜、线粒体、内质网以及肝脏组织中的部分脂滴周围及微绒毛中AKP依次出现在各组织中的细胞膜、细胞核、溶酶体、内质网以及肝脏组织中的脂滴周围及微绒毛中。

The oversaturation of cholesterol in bile results in the formation of cholesterol stones. The liver is the organ which synthesizes cholesterol and secretes bile. Liver lipid metabolism research helps to explain the mechanism of cholelithiasis. Utilizing biochemical and biomolecular technology, we have done experimental work as follows: Analyse the ingredient changes of blood and bile samples from patients.

肝脏是合成胆固醇、分泌胆汁酸的器官,对肝脏胆固醇代谢的研究有助于阐明胆石症形成的原因,本文用生物化学和分子生物学的方法,分析了病人血液、胆汁中各种脂类成份的变化,以及肝脏胆固醇合成限速酶3-羟-3-甲戊二酸辅酶A还原酶(HMG-CoA Reductase)、胆汁酸合成限速酶7α-羟化酶(7α-hydrolase)的基因表达。

The effects of different concentration of Hg2+,Cr3+ and Pb2+ on the membrane peroxidation content and the activities of antioxidant enzymessuperoxide SOD, POD, CAT in mung bean in podding period were studied.

研究了不同浓度Hg2+、Cr3+和Pb2+胁迫条件下,绿豆花荚期叶片中膜脂过氧化物含量及抗氧化酶,包括超氧化物歧化酶(SOD、过氧化物酶和过氧化氢酶活性的变化情况。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法,从人组织细胞总RNA中扩增可溶性TWEAK胞外区(sTWEAK1和sTWEAK2)的cDNA序列及全长编码序列,用琼脂糖凝胶电泳分析PCR产物,胶回收目的基因片段,连接到pMD18-T克隆载体中,转化大肠杆菌K802,PCR和酶切筛选阳性克隆,全自动DNA测序验证序列;(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中,分别转化大肠杆菌BL21和TB1,PCR筛选和酶切鉴定,阳性克隆用IPTG诱导表达,表达产物用SDS-PAGE分析和Western blot验证融合蛋白;(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白;(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测,贴壁细胞用结晶紫染色法,悬浮细胞用磺酰罗丹明B染色法,酶标仪检测OD值;(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量;(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况;(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响;(8)用双荧光素酶报告基因检测法,测定表达产物对敏感细胞NF-κB的影响;(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上,用脂质体转染法转染HEK293细胞,PCR鉴定重组质粒。

In present study, we observed effect of taurine on liver lipid metabolism in rat, an animal capable of synthesizing substantial amount of taurine and extensively used in studies on lipid metabolism, and tried to explain its biochemical basis by investigating several hepatic microsomal membrane-bound enzymes activities related to lipid metabolism.

本实验中,我们将观察牛磺酸对大鼠肝脂质代谢的影响,并观察牛磺酸对大鼠肝脏微粒体脂质代谢酶活力的影响,以探讨牛磺酸影响肝脏脂质代谢的可能机制。

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