组氨酸

Shandong sinder technology limited company,qingdao 266061,shandong,chinaabstract:to provide the eukaryotic expression vector carrying 6x histidine-tag and human growth hormone in expremental animal and cell culture,according to the published nucleotide sequence of the lactofericin b and human growth hormone,synthesis sequence was ligated with pmt-18 vector.

以牛乳素为目的基因，构建含有组氨酸标签、人生长激素信号肽的真核表达载体，为利用牛乳素在活体动物及培养细胞中的表达提供基因材料。

To prepare recombinant fox growth hormone, we amplified its cDNA from silver fox pituitary tissue by RT-PCR and cloned into yeast shuttle vector pPIC9K down stream of α-factor signal peptide sequence by SnaB I and Not I restriction sites. The recombinant secretion vector pPIC9K/fGH, linearized by Sal I, was transformed into histidine-deficient Pichia pastoris strain GS115 by electroporation.

为制备重组狐狸生长激素，采用RT-PCR方法，从银狐垂体中扩增fGH cDNA基因，利用SnaB I和Not I位点将fGH基因插入到酵母分泌型表达载体pPIC9K中α-因子信号肽的下游，构建成fGH基因的酵母分泌型表达载体pPIC9K/fGH，载体经Sal I酶切线性化后，通过电转移将线性化的pPIC9K/fGH转化到组氨酸缺陷型酵母宿主菌GS115中。

As for the ester exchange reaction on phosphorus, it was found that N-phosphoryl histidine the fastest one among all of the N-phosphoryl aminoacids. The results studied by MNDO method indicate that due to theparticipation of imidazole group, the formation of hexa-coordinatephosphorus intermediate made the iso-propoxyl which is opposite toimidazole group more active than the other ones and much easier to leave.When iso-propoxyl left, a new penta-coordinate phosphorus intermediate wasformed, and it is sterically favorable to be attacked by an alcohol from theopposite direction of imidazole.

对于磷上酯交换反应，实验中发现N-磷酰组氨酸反应速度明显高于其它N-磷酰氨基酸，理论研究表明，侧键咪唑基参与下六配位磷中间体形成后，位于咪唑基对面的异丙氧基反应活性大大提高，即从咪唑基的对面位置发生异丙氧基的离去和另一分子醇的进攻，在能量上和空间上都是有利的。N-磷酰丝氨酸的磷上酯交换反应有着类似的特点。

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。

A pair of primers were designed based on M1 gene sequence of known H5N1 Avian influenza virus.M1 gene was cloned from total RNA,extracted from tissue of H5N1 subtype virus inoculated embryo by reverse transcriptase-polymerase chain reaction using high proofreading polymerase(Pyobest~TM DNA Polymerase),and expressed using Invitrogen champion~TM pET directional TOPO expression system.Recombinant protein containing polyhistidine(6xHis) tag in N-terminal about 29.8kDa in size,wac obtained and purified.

根据已发表的禽流感病毒M1基因序列设计合成PCR克隆引物，自接种H5N1亚型病毒的鸡胚组织中提取RNA，反转录后采用高可信度DNA聚合酶经PCR扩增M1基因，采用Invitrogen定向表达系统(ChampionTMpET directional TOPO expression system)进行克隆表达，纯化获得N末端携带多聚组氨酸标签的重组蛋白，分子量约29.8 kDa。

The effects of different amino acids on the morphologies and orientations of electrodeposited ZnO were studied.

考察了不同种类的氨基酸（组氨酸，赖氨酸，氨基乙酸，谷氨酸）对电沉积制备的ZnO的形貌及晶体取向的影响。

Objective To investigate the changes in the fragile histidine triad gene expression in esophageal aqua moos cell carcinomas and the possible mechanism.

目的 探讨脆性组氨酸二联体基因在食管鳞癌中表达的变化及可能机制。

Coli JM109 (DE3) transformant containing the 〓-tagging vector with the gene leuS was approximately 110 times that of JM109 (DE3)(the host strain without the vector). The overproduced 〓-fusion leucyl-tRNA synthetase can be purified to homogeneity under native condition within two hours by one step affinity chromatography with an overall yield of 55%.

高表达的N—端带有6个连续组氨酸的LeuRS经过一步Ni-NTASuperflow亲和层析可以达到90％以上的纯度，产率为55％，纯化时间仅为2小时，而且LeuRS所带的〓-标签对酶的结构和功能都没有影响。

Objective: The purpose of this study was to determine whether potentiating induction of apoptosis in cervical cancer cell line SiHa lacking full-length FHIT transcripts and endogenous FHIT protein by cisplatin via introduction of extopic FHIT gene. Meanwhile, to investigate if the apoptosis of the SiHa cells is induced by FHIT gene.

目的 验证外源性脆性组氨酸三联体抑癌基因的导入是否诱导宫颈癌SiHa细胞凋亡，观察FHIT导入与顺铂联合应用诱导宫颈癌SiHa细胞凋亡的作用是否有协同效应，为联合基因治疗和化疗药物治疗人宫颈癌提供实验依据。

Photosensitizer rose bengal reacts with O 2 to generate singlet oxygen ( 1O 2) under illumination .

光敏剂RB在光照射下与O2反应产生1O2，1O2与组氨酸或咪唑反应的中间产物使RNO发生氧化，导致RNO在440nm处吸光度减小，此即为RNO脱色反应。

However, as the name（read－only memory）implies, CD disks cannot be written onorchanged in any way.

然而，正如其名字所指出的那样，CD盘不能写，也不能用任何方式改变其内容。

Galvanizes steel pallet is mainly export which suits standard packing of European Union, the North America. galvanizes steel pallet is suitable to heavy rack. Pallet surface can design plate type, corrugated and the gap form, satisfies the different requirements.

镀锌钢托盘多用于出口，替代木托盘，免薰蒸，符合欧盟、北美各国对出口货物包装材料的法令要求；喷涂钢托盘适用于重载上货架之用，托盘表面根据需要制作成平板状、波纹状及间隔形式，满足不同的使用要求。