Tn5 and Sleeping Beauty transpositional transgenesis by direct cytoplasmic injection in bovine zygotes.

<p>A,D) Blastocysts produced by injection with (ME)<i>egfp</i>:Tn5 transposome. B,E) Blastocysts produced by injection with pT2/Venus and pCMV-SB100X. C,F) Blastocysts produced by injection with pT2/Venus alone. A-C) Under bright light. D-F) Under blue light (488 nm). G) Embryo Southern blot performed on DNA amplified from Tn5 and Sleeping Beauty system injected blastocysts. WGA was performed to obtain enough DNA for Southern blot. Lane 1: 10 pg <i>Sal</i>I-<i>Xba</i>I pCX-EGFP 1.6 Kb positive control. Lane 2: 1 ng <i>Sal</i>I-<i>Xba</i>I pCX-EGFP 1.6 Kb positive control. Lane 3,5: Empty lanes. Lane 4: IVF blastocyst (negative control). Lane 6–9: Tn5 transposon control. Lane 10–13: Tn5 transpositional transgenesis blastocysts. Lane 14–17: Sleeping Beauty transpositional transgenesis blastocysts. MW: NEB 1 kB Ladder of molecular weight.