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The full-leng coding sequence of ALK1 was cloned into pcDNA3.l+using standard protocols. The constructed pcDNA3.1+ALK1 plasmid were transfected into HUVECs. The proliferation and migration of HUVECs were detected by boyden champer and flow cytometry.

脂质体转染pcDNA3.1+ALK1到HUVECs，流式细胞仪检测HUVECs增殖的改变，boyden小室检测ALK1对HUVECs迁徙的影响。

An infectious clone of the sat-RNA of BBSV-X was also constructed by placing the cDNA under the control of the 35S promoter. This plasmid could infect Chenopodium amaranticolor with the help of in vitro transcripts of BBSV-X.

构建了35S启动子控制下的BBSV卫星RNA全长侵染性克隆，侵染性检测表明，所获得的3个克隆均有较好的侵染性。

Objective: To construct and identify the chimeric MS/hIL12 eukaryotic expression plasmid.

目的：克隆含信号肽的Mtb8.4/hIL12嵌合基因，构建其真核表达质粒，并进行鉴定。

Methodological design, synthesis is aimed at anoxic and revulsive factor - the nucleotide of few of particularity short cable length of 1 α is acerbity, compose builds anoxic and revulsive factor - of ShR-NA of 1 α particularity recombine plasmid, stability turns catch; of cell of colonic cancer SW 480 to use chloridize cobalt preparation is anoxic and revulsive education radical, imitate tumour is anoxic condition.

方法设计、合成针对缺氧诱导因子-1α的特异性短链寡核苷酸，构建缺氧诱导因子-1α特异性shR-NA的重组质粒，稳定转染结肠癌SW 480细胞；采用氯化钴制备缺氧诱导培养基，模拟肿瘤缺氧状态。

Methods Trizol reagent from clamworm digestive tract epithelial cells extracted methods Total RNA. The mRNA was purified and reverse transcripted into cDNA. The cDNA was linked into plasmid pDNR-Lib after digested by nucleate endonuclease Sfi I to construct cDNA library.

分离沙蚕消化道上皮细胞，使用Trizol试剂提取总RNA，纯化mRNA，用MMLV逆转录成cDNA，用Sfi I核酸内切酶酶切后连接到pDNR-Lib质粒中，构建cDNA文库。

YIp5 plasmid which is replicable in E. coli but not in yeast is served as clonic carrier and ARS sequence in total DNA of rice cloned by air gun experiment.

以能在大肠杆菌复制，但不能在酵母菌中复制的YIp5质粒作为克隆载体，鸟枪法克隆了水稻总DNA中的ARS顺序。

Plasmid DNA profiles analysis offer scientific reference to molecular epidemiology investigation of avian colibacillosis.

质粒DNA图谱分析为从分子水平上进行鸡大肠杆菌病流行病学调查提供了基础材料。

Then we used Arabidopsis CaM2 isoform as bait to screen the two-hybrid library. The combinant bait plasmid pGBKT7-ACaM2 was constructed.

其次，以拟南芥2号亚型钙调素(ACaM2)为诱饵蛋白对双杂交文库进行了筛选。

Methods The coding gene of Sj32KD antigen was amplified from the cDNA library of the adult parasite by PCR and cloned into the prokaryotic expression vector pET28a to construct a re- combinant plasmid Sj32-pET28a.

应用PCR方法克隆Sj32KD抗原基因，并在大肠杆菌系统中进行表达，利用His亲和层析方法纯化融合蛋白，并应用重组的Sj32KD抗原检测羊日本血吸虫病。

Then the combinant plasmid was conducted into ACC-2 cell by electroporation. ACC-2 cells stably expressing CD was obtained by 10-day positive selection with 400 μg/mL G418. Total RNA was extracted and the expression of the CD gene in transfected ACC-2 cells was identified by RT-PCR. RESULTS: PCR yielded a fragment of l280bp and CD was verified by sequence analysis.

测序正确后，将其亚克隆到质粒表达载体pIRES中，构建以内部核糖体进入位点相连的CD基因的质粒表达载体pIRES-CD，采用电穿孔法，以质粒表达载体转染ACC-2细胞，用400μg/mL的G418筛选10d，获得稳定表达CD基因的ACC-2细胞系，提取该细胞的总RNA，用RT-PCR检测CD基因的表达。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！