[3] Restriction site bank vectors for cloning in gram-negative bacteria and yeast

摘要: Publisher Summary This chapter describes the application of restriction site banks for construction Escherichia coli vectors, wide-host-range cosmid vectors (able to grow in most gram-negative bacteria), and E. –yeast shuttle vectors. The summarizes bank series that have been constructed as tools genetic manipulation DNA. advantage is they maximize choice enzymes can be used cloning facilitate subsequent subcloning deletion analysis. Thus, permit, a single plasmid, variety operations could otherwise done only with combination several different vector systems. Of known endonucleases recognize 6-bp palindromes, 39 unique recognition sites pJRD184 four others, while not unique, also without disturbing essential functions. example illustrated involves genes vanillate utilization from an uncharacterized pseudomonad ATCC 19151. Vanillate key intermediate degradation lignin molecule, one world's abundant carbon sources potential source industrial raw material energy. system useful demonstrate generality cloning, conjugal transfer, complementation method gene isolation.