Map of lentiviral vector.

<p>Map of lentiviral vector.</p

RT-PCR amplification of a specific fragment of the lentiviral vector after incubation with sperm cells.

<p>Lane M: DNA Ladder DL2000 (2000, 1000, 750, 500, 250 and 100 bp from top to bottom). Lane 1: Negative control with H₂O as template. Lane 2: Negative control with total RNA extracted from sperm alone as template, both of which showed no PCR products. Lanes 3–6: Specific PCR products were detected in sperm samples incubated with lentiviral particles for 2, 4, 6 and 8 h, respectively.</p

Absorption of lentiviral particles by pig spermatozoa under a scanning electronic microscope and immunochemical detection of lentiviral particles on spermatozoa.

<p>A: lentiviral particles (40000×). B: Two lentiviral particles are clearly attached on the head of one spermatozoa (10000×). C: Two lentiviral particles absorbed on a spermatozoa tail (25000×). D: Observation of interaction between a lentiviral particle and the spermatozoa surface in an enlarged image (60000×). E: A lentiviral particle entering a spermatozoon (7500×). Immunuchemical detection of lentiviral particles attached on sperm was performed using a monoclonal mouse anti-VSV-Glycoprotein antibody, F was view field of porcine spermatozoa (200×), and red fluorescence was clearly observed emitted at 570 nm (G, 200×).</p

Identification of transgenic pigs by Southern blot analysis.

<p>Southern blot was performed under optimized condition with a Dig-lableled probe of 649 bp fragment. Samples in lanes 1 and 2 are from PCR-positive the founder No. 18 and No. 41, respectively, lanes 3–8 are their offsprings; The specific hybridization signal was detected in Lanes 1, 2, 4, 9; lanes 3, 5, 6, 7, 8 are negative, respectively; lane 10 is wild type control; lane 11 is positive controls (The plasmid pLV-siRNA); M is DNA Molecular Weight VII (DIG-labeled Roche).</p

PCR amplification of transgene DNA sequences in piglets.

<p>PCR amplification of transgene DNA sequences in piglets.</p

Number of lentiviral particles associated with sperm cells after incubation (n = 3).

<p>Number of lentiviral particles associated with sperm cells after incubation (n = 3).</p