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    • 47. 发明授权
    • Plasmid cloning vector pAS1
    • 质粒克隆载体pAS1
    • US4925799A
    • 1990-05-15
    • US819406
    • 1986-01-16
    • Martin Rosenberg
    • Martin Rosenberg
    • C12N15/73
    • C12N15/73
    • A plasmid cloning vector containing both transcriptional and translational regulatory sequences derived from the bacteriophage lambda genome was constructed to achieve high level expression of prokaryotic and eukaryotic genes. The system utilizes a plasmid vehicle carrying the strong, regulatable lambda promoter, P.sub.L, and host lysogens into which this vector can be stabily transformed. The lysogen synthesizes sufficient repressor (cI) to control P.sub.L expression and thereby stabilize plasmids which carry such a highly efficient promoter. Use of a temperature sensitive repressor permits simple, rapid induction of P.sub.L transcripts at any given time. Efficient transcription of essentially any coding sequence is assured by providing the phage lambda antitermination factor, N, and a site on the transcription unit for its utilization (Nut site). This pAS1 plasmic closely resembles the earlier constructed pKC30cII, also a regulatory protein which activates promoters for lysogenic development.
    • 构建含有衍生自噬菌体λ基因组的转录和翻译调控序列的质粒克隆载体,以实现原核和真核基因的高水平表达。 该系统使用携带强的可调节的λ启动子,PL和宿主溶菌体的质粒载体,其中该载体可以稳定地转化到其中。 溶菌酶合成足够的阻遏物(cI)以控制PL表达,从而稳定携带这种高效启动子的质粒。 使用温度敏感的阻遏物允许在任何给定时间内简单,快速地诱导PL成誊本。 通过提供噬菌体λ抗突变因子N和转录单位上用于其利用的位点(Nut位点)来确保基本上任何编码序列的有效转录。 该pAS1等位基因非常类似于早期构建的pKC30cII,也是一种调节蛋白,其激活用于溶原性发育的启动子。