- corotation
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- After obvious cytopathogenic effects developed , virus - contained supematants were harvested , and the progeny viruses were screened for lacz - expressing viruses by a plaque assay using x - gal . single blue plaques were picked , and a recombinant prv stably expressing lacz gene ( designated as rprv - lacz ) was obtained after ten cycles of plague purification and pcr identification . the results showed that the lacz gene expression cassette was stably expressed in the recombinant rprv - lacz derived from bartha - k61 strain
该载体与具有高度感染性的bartha - k61株基因组dna通过脂质体加plus法共转染vero细胞，采用甲基纤维素固定病变， x - gal染色，经过10代蓝斑纯化获得了一株稳定表达lacz基因的ge tk基因缺失突变株，命名为rprv - lacz 。 - Fusion gene by pcr was inserted into bombyx mori baculovirus transfer vector pbacpak . 8 and contransfected with lineared dna of bm - bacpak6 virus into bmn cells . the homologous recombination occurred inside the cells , and the recombinant virus bacpak - 6aa - hgm - csf was expressed , as identified by pcr and southern hybridization . the bmn cells and the fifth instars were infected by the recombinant virus bacpak - 6aa - hgm - csf
本研究首先通过pcr将家蚕杆状病毒多角体蛋白起始密码子后的18个碱基引入到hgm - csf基因的5 ’端之前，然后将融合基因重组与家蚕杆状病毒转移载体pbacpak8中，获得重组转移载体pbacpak8 - 6aa - hgm - csf ，并与线性化bm - bacpak6dna共转染家蚕细胞株，获得重组病毒bacpak - 6aa - hgm - csf 。 - The pbacph - egf plasmid dna was used to co - transfect bmn cell with the modified bombyx mori baculovirus bm - bacpak dna , which was first linearized by aocl . after two rounds of plaque isolation , the recombinant virus ( named bmbacph - egf ) was further verified with pcr and dot hybridization . the recombinant bmbacph - egf virus was used to infect bmn culture cells at a moi of 10
重组转移载体dna与经bsu36i酶切线性化的修饰型病毒bm - bacpakdna共转染家蚕bmn细胞，两轮空斑筛选和纯化，获得重组病毒rbmbacph - egf ，经pcr扩增、 dna点杂交等方法鉴定证实重组病毒中已正确插入ph - egf融合基因。 - Angiostatin ( kl - 3 ) gene was inserted into bombyx mori baculovims transfer vector pbacpaks and cotransfected with lineared dna of bm - bacpak6 virus into bmn cells . the homologous recombination occurred inside the cells , and the recombinant virus bacpak - angiostatin was expressed , as identified by pcr and dna dot blotting
本研究首先将人血管抑素( angiostatin )基因重组于家蚕杆状病毒转移载体pbacpak8中，获得重组转移载体pbacpak - angiostatin ，并与线性化病毒bm - bacpak6dna共转染家蚕细胞株，获得重组病毒bacpak - angiostafin 。 - On the other hand , our previous studies have shown that the nr2b construct with a gfp tag in its n - terminus cotransfected with nr1 subunit can reach the cell surface and be detected by surface staining with anti - gfp antibody in live cells . the nr2b construct will be retained in the endoplasmic reticulum ( er ) when expressed alone in heterologous cells
我们以往的研究表明，用绿荧光蛋白( gf内在信号肤下游n末端标记nrzb亚单位，与nri共转染时能形成与野生型相似的nmda受体通道，且可以通过抗gfp抗体标记活细胞膜表面表达的nmda受体复合物。