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The recombination vector was digested by Pac I enzyme and transfected into 293A cell by Lipofectamine method to obtain recomb...

该载体经PacI酶切，脂质体法转染人胚肾293A细胞，获得重组腺病毒载体pAD-E7.pAD-E7转染人HaCaT细胞，激光共聚焦显微镜分析E7蛋白表达情况。

Two relevant sites of enzymatic digestion were added to the mTNF-α by PCR. The mTNF-α was linked to the 3'end of m/〓 in pGEX4T-1 vector. The prokaryotic expression vector pGEX4T-1m/〓-mTNF-α was constructed successfully. After induction and expression by IPTG, the expression of two kinds of fusion protein is 15% and 12% of total bacteria proteins respectively. The anti-HCC bifunctional antibodies m/〓-mTNF-α were identified by electrophoresis after the inclusion bodies were purified, denature, renature, re-purified, digested by thrombin and further purified.

采用PCR的方法在mTNF-α的两端加上所需要的酶切位点，将之连接在m／〓的3'端，构建原核表达载体pGEX4T-1 m／〓-mTNF-α，通过IPTG的诱导表达之后，两种融合蛋白的表达量分别占细菌总蛋白的15％、12％，表达产物经包涵体的纯化→变性→复性→纯化→凝血酶酶切→进一步纯化后，可以得到纯度为电泳纯的m／〓-mTNF-α抗肝癌双功能抗体。

The sample was digested by microwave,magnesium,zinc,iron in the Silurus lanzhouensis Chen were determined using flame atomic absorption spectrometry.

使用微波消解处理样品，火焰原子吸收光谱法测定了兰州鲶中的铁、锌、镁含量。

METHODS: Fetal mouse liver was sterilely obtained, combed using cell scrape, and digested in type IV collagenase.

无菌操作取出胎鼠肝脏，用细胞刮梳理后用IV型胶原酶消化。

The serum selenium is first digested by microwave treatment and then chemically purified before analysis by using sulphydryl cotton. The optimum conditions were found to b as follows:(1) The sulphydryl cotton column is prewashed with 5 mol/L hydrochloric acid;(2) Se is desorbed by adding 1 mL of concentrated nitric acid and (3) the optimum amount of sulphydryl cotton is 0.14 g. This preconcentration step is required in order to remove matrix-interfering elements, such as Na, Mg, K and Ca.

采用微波消解－巯基棉分离对人血清样品进行前处理，通过对巯基棉分离血清中硒的条件优化，结果表明，柱条件为5mol/L HCl、解脱Se条件为1mL重蒸HNO3、巯基棉最佳用量为0.14g时，能够有效分离出人血清样品中的Na、Mg、K、Ca等碱金属和碱土金属，同时硒的回收率最高，达到92%以上。

Lastly, specimens contain HPV16 DNA were digested with restricted enyzme BamHI, and done with southern transprint to hybridize with digoxigenin-labeled probe of HPV16 genome through nuclear acid hybridization. Results (1)Among 60 cervial carcinoma specimens,the HPV16 E5 positive rate is 33%(20/60) by PCR, 70%(42/60)、58%（35/60）by nuclear acid dot hybridization with HPV16 E5 probe and HPV16 probe .

结果（1）PCR法 60例宫颈癌组织中HPV16 E5 片段的阳性率为33%（20/60）；（2）斑点杂交法60例宫颈癌组织中HPV16 E5阳性率为70%(42/60)，HPV16阳性率为58%（35/60）；（3）宫颈癌组织中HPV16 E5基因的突变率为40%（8/20），迁移位置不甚一致，随机抽取一例E5变异的PCR阳性产物进行测序分析，结果发现在E5片段中有2处变异， nt3979A→C颠换，nt3988T→C转换，导致E5编码区出现1个氨基酸的有义变异。

Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal antibody.

参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNABac-N-Blue~(TM DNA共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。

Methods Affinity chromatography purified plasminogen was digested by pancreatic elastase and angiostatin was obtained by purifying the digestion with affinity chromatography and dialysis. T739 mice were inoculated with LA795 cells. Angiostatin was given to part of inoculated and uninoculated mice by intraperitoneal injection fourteen days after the inoculation. The treatment lasted 20 days and the size of tumor, survival period, behavior of the mice and pathology of lungs, livers and kidneys were observed.

用亲和层析方法得到的血浆纤维溶酶原经胰蛋白酶消化、层析柱亲和层析、透析，得到纯化的血管抑素；以LA795细胞接种于T739小鼠；接种后第15日起腹腔内注射血管抑素共20 d，观察动物行为，测量荷瘤鼠肿块直径，记录生存期，另设未治疗对照组及观察血管抑素副作用对照组，行肺、肝、肾脏病理学检查。

Methods utricular sensory epithelia of postnatal day 1 rats were isolated and digested by thermolysin, then were transferred to a solution containing 0.125% trypsin and 0.125% collagenase for 8 min at 37℃ to harvest the pure utricular sensory epithelial cells.

取出生1天（ postnatal day 1， p1 ）大鼠的椭圆囊感觉上皮，嗜热菌蛋白酶消化处理，获得纯椭圆囊感觉上皮，再采用消化法进行原代培养，培养基为dmem。

The fresh dewater digested sludge of municipal wastewater treatment plant was suited for vaccinal sludge at the startup of anaerobic reactor and the concentration should not be less 10mg/L; the COD of feed wastewater should be lower than 5000mg/L, and VOL was 20 percent of total VOL; the alkalinity of effluent should be more than 3000mg/L; on the condition of the COD removed 80% can the VOL be elevated.

反应器启动时接种污泥可以选用城市污水处理厂新鲜的脱水消化污泥，浓度不低于10mg/L；进水有机物浓度不高于5000mg/L，容积负荷为满负荷的20％；出水碱度要高于3000mg/L，温度稳定；在有机物的降解率达到80％后再提高负荷。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。