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Bacterial plasmid vectors for cloning foreign DNA, analysis of transcription signals, and expression of fusion proteins, etc
Bacterial plasmid vectors for cloning foreign DNA, analysis of transcription signals, and expression of fusion proteins, etc
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机译：用于克隆外源DNA，分析转录信号以及表达融合蛋白的细菌质粒载体
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摘要
Bacterial plasmid vectors based on alpha -amylase, are new. Bacterial plasmid vectors based on alpha -amylase, with a wide host spectrum and multiple uses, particularly amongst gram-positive bacteria, may support: cloning of foreign DNA with positive selection; isolation of any constitutive promoter; isolation of inducible/repressible promoters, where a single regulatory agent is known; quantitative and qualitative analysis of promoter activities, operator sequence, ribosome binding sites, start codons, signal peptides or display signals for cell wall proteins; constitutive expression of foreign proteins; constitutive expression of fusion proteins containing alpha -amylase, where it is possible to choose, whether the product is cytoplasmic or extracellular, and whether the fusion protein will be at the 5' or 3' end; cell surface display of fusion proteins in gram-positive bacteria; alpha -amylase activity test without cell information; alpha -amylase activity test in organisms with their own alpha -amylase, where this is not heat stable; purification of fusion proteins with strong adsorption; detection of alpha -amylase, or fusion derivatives, through color activity in a native or denatured polyacrylamide gel; availability for Escherichia coli with two alternative replication origins and two alternative resistance markers; and insertion in other bacteria through three alternate methods. Independent claims are also included for: pre-made medium that makes it possible for positive selection of alpha -amylase activity without use of I2/I- solution; a method of cloning with help of a pre-linearized vector to faultlessly construct a shuttle-vector in accordance with (m) for the insertion of vectors in gram-positive bacteria, choice between three differing copy numbers in target organisms; a helper vector and degenerate oligonucleotide for the possible direct use amongst vectors from above in gram-positive bacteria; and vector and selection method to isolate regulated promoters in accordance with 1.c) by selective cell sorting.
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