克隆

PAP-C23, amplified from mature pokeweed antiviral protein gene by PCR, was cloned into the expression vector PET101, and then the recombinant plasmid was transformed into Escherichia coli BL21(DE3). The recombinant mutant PAP was expressed high-levelly induced with IPTG.

用KCL溶液染SDS-PAGE胶，切下表达的目的蛋白，冰浴研磨后用磷酸缓冲液溶解，再加入等体积的福氏不完全佐剂，乳化完全后，免疫家兔，制备抗PAP蛋白的多克隆抗体，间接ELISA法测定所制备的抗血清的效价为1：1000。

Transgenic tobacco plants were obtained through screening with kanamycin. The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants. Pokeweed antiviral protein Ⅱ is expressed with high level in summer leaves. The expression of PAPⅡ is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPⅡ gene by RT-PCR and then the gene was cloned into E. coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPⅡ gene were then transferred into E. coli strain BL21 (DE3)-plysS and Pachia pastoris GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

The transgenic tobacco plants could delay TMV infection for about 25 days compared with non-transgenic tobacco plants.Pokeweed antiviral protein II is expressed with high level in summer leaves. The expression of PAPII is regulated by season. The total RNA was extracted from pokeweed (Phytolacca americana L.) leaves in summer using the method of TRIzol and used as template to amplify the PAPII gene by RT-PCR and then the gene was cloned into E.coli expression vector and secreted expression pPIC9K vector. The two vectors with PAPII gene were then transferred into E.coli strain BL21 (DE3)-plysS and Pachia pastor is GS115 strain respectively. The specific protein was produced induced by IPTG and methanol.

由于美洲商陆抗病毒蛋白Ⅱ是一个受季节调控表达的蛋白，本实验以美洲商陆夏季叶片为材料，通过对其总RNA的提取、反转录、并用PAPⅡ的特异引物进行PCR扩增，对PAPⅡ进行了基因克隆、序列分析，结果扩增出来的PAPⅡ基因与已经报道的序列同源性是99.9％，然后将该基因构建到原核、真核表达载体上，分别转化了大肠杆菌和毕赤酵母并对它们进行了诱导表达，在两个表达系统中均获得了有活性的PAPⅡ表达蛋白，体外生物测定表明表达的蛋白均具有抑制病毒的活性，PAPⅡ基因在酵母中还没有表达的报道，这为获得具活性PAPⅡ蛋白提供了一种简便可行的方法。

On May 6 the electric potential is the island 圣皮埃尔 harmonious clone to the Canadian northeast seacoast law the inhabitants and France takes the lead in the Americas each place alien and the overseas inhabitant to go to the polling booth, started French presidents the election second turn of voting.

5月6日电位于加拿大东北海岸的法属小岛圣皮埃尔和密克隆的居民们与法国在美洲各地的侨民和海外居民率先前往投票站，开始了法国总统大选第二轮投票。

Cleavage polyembryony The splitting of an embryo into several identical parts, each of which may develop into a mature embryo. It is form of natural cloning.

裂生多胚现象：一个受精卵在发育过程中胚原细胞分裂为几个胚的现象，是自然克隆的一种形式。

Results］ Two fragments of 1 258 bp were amplified from Polygonum multiflorum.

结果］ 从何首乌叶cDNA中克隆出了长度为1 258 bp的基因片段。

But thefunctions of SOD-like genes were not approved.We cloned SOD gene fromBombyx mori Nuclear Polyhedrosis Virus genome by PCR.

我们克隆了家蚕杆状病毒的sod基因，测定其核苷酸序列与人源的SOD1基因同源性为56％，其中铜、锌离子的结合位点和酶活性中心位点相当保守。

Although Americans have been much less bothered about biotechnology and food—they couldn't seem to care less about eating GM crops—it's safe to say that few people are thrilled by the prospect of putting a fork into a cloned porterhouse.

尽管美国人对于生物技术和食品的担心没那么多，但是，他们对食用转基因农产品的关注似乎一点也不少。可以有把握地说，没有什么人会对食用克隆牛排胆战心惊。

All these studies will potentate the progress in field of study on pathogenesis and new therapies.

因此，从基因的克隆化角度入手，也是阐明病毒性肝炎发

In order to understand the mechanisms underlying powdery mildew resistance of wheat, its necessary to isolate genes associated with the resistance reactions.

为了了解小麦抗白粉病的机制，本文开展了对小麦抗白粉病相关基因的克隆研究。

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。