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不同条件下人乳白蛋白YAC的高效定点打靶修饰
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作者 张绪峰 吴国祥 +2 位作者 朱克军 焦炳华 成国祥 《第二军医大学学报》 CAS CSCD 北大核心 2004年第6期633-637,共5页
目的?对人乳白蛋白酵母人工染色体(HLA-YAC)进行定点改造,以便进一步利用该载体作与基因表达调控相关的 研究。方法:借助kar1突变使YPH925菌株与异交配型菌株交配时丧失核融合的能力,把HLA-YAC由酵母AB1380转移到 his3缺陷的YPH925菌株... 目的?对人乳白蛋白酵母人工染色体(HLA-YAC)进行定点改造,以便进一步利用该载体作与基因表达调控相关的 研究。方法:借助kar1突变使YPH925菌株与异交配型菌株交配时丧失核融合的能力,把HLA-YAC由酵母AB1380转移到 his3缺陷的YPH925菌株。分别以HIS3(对YPH925菌株)和G418R(对YPH925和AB1380两种菌株)为选择标记,用含 HLA-YAC的酵母基因组为模板,PCR克隆人乳白蛋白(HLA)基因的5′和3′端非翻译区各684bp和940bp为同源臂,构建 打靶载体,线性化的打靶载体经电转化或醋酸锂转化转入酵母内,以菌落PCR为鉴定方法,辅以测序验证。结果:HLA-YA 在目的位置完成了定点打靶修饰,G418R的筛选效率高于HIS3,醋酸锂转化方法的打靶效率稍高于电转化。结论:对YA 的成功修饰为在动物体内的表达及调控研究打下了基础。 展开更多
关键词 人乳白蛋白 YAC 高效定点打靶修饰 酵母人工染色体 酵母人工染色庠转移
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Binding interactions of pefloxacin mesylate with bovine lactoferrin and human serum albumin 被引量:2
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作者 FAN Ji-cai CHEN Xiang WANG Yun FAN Cheng-ping SHANG Zhi-cai 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2006年第6期452-458,共7页
The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant ... The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant K and the binding sites n were obtained by fluorescence quenching method. The binding distance r and energy-transfer efficiency E between pefloxacin mesylate and bovine lactoferrin as well as human serum albumin were also obtained according to the mechanism of Forster-type dipole-dipole nonradiative energy-transfer. The effects of pefloxacin mesylate on the conformations of bovine lactoferrin and human serum albumin were also analyzed using synchronous fluorescence spectroscopy. 展开更多
关键词 Pefloxacin mesylate (PFLX) Bovine lactoferrin (BLf) Human serum albumin (HSA) Fluorescence spectra Energy-transfer efficiency
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