000			01669nam a2200181Ia 4500
003			MX-MdCICY
005			20250625153930.0
040			_cCICY
090			_aB-13832
245	1	0	_aIn vitro insertional mutagenesis with a selectable DNA fragment
490	0		_vGene, 29(3), p.303-313, 1984
520	3		_aA new method for in vitro insertional mutagenesis of genes cloned in Escherichiu coli is presented. This simple procedure combines the advantages of in vitro DNA linker mutagenesis with those of in vivo transposition mutagenesis. It makes use of the D fragment, a 2.0-kb DNA segment consisting of an antibiotic resistance gene (the Sm'/Spc' gene of the R1OO.l plasmid)flanked by short inverted repeats carrying transcription and translation termination signals and synthetic polylinkers. The Sz fragment is inserted into a linearized plasmid by in vitro ligation, and the recombinant DNA molecules are selected by their resistance to streptomycin and spectinomycin. The D fragment terminates RNA and protein synthesis prematurely, thus allowing the definition and mapping of both transcription and translation units. Because of the symmetrical structure of Sz, the same effect is obtained with insertions in either orientation. The antibiotic resistance gene can be subsequently excised from the mutated molecules, leaving behind its flanking restriction site(s).
700	1	2	_aPrentki, Pierre
700	1	2	_aKrisch, Henry M.
856	4	0	_uhttps://drive.google.com/file/d/1d7jZAYGRLPN8hZQXKEuvwjyx9Ve_x69z/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
942			_2Loc _cREF1
008			250602s9999 xx |||||s2 |||| ||und|d
999			_c48029 _d48029