Primers and PCR conditions used for the detection of target genes in laying hens digesta, blood, tissues, and eggs samples.

1<p>PCR conditions for <i>ivr</i> F & R included 1 cycle at 95°C for 5 min, 40 cycles of 94°C for 30 s, down to 61.5°C for 45 s and back up to 72°C for 45 s and 1 cycle of 72°C for 10 min; <i>phy</i>A2-F & R; 1 cycle at 94°C for 5 min, 40 cycles of 94°C for 30 s, down to 61.5°C for 45 s and back up to 72°C for 45 s and 1 cycle of 72°C for 10 min; <i>ov</i>-F & R; 1 cycle at 94°C for 3 min, 35 cycles of 94°C for 30 s, down to 55°C for 30 s and back up to 72°C for 45 s and 1 cycle of 72°C for 10 min.</p

Western blot detection of the <i>phy</i>A2 protein in corn, diet, digesta, blood, tissues, and eggs of laying hens fed diets containing control corn (CC) and phytase transgenic corn (PTC) for 16 weeks¹.

1<p>Number of samples that tested positive for the <i>phy</i>A2 protein out of 8 samples analyzed. One sample was tested per laying hen (n = 8 laying hens per treatment).</p>2<p>Percentage of samples positive for the <i>phy</i>A2 protein taken from PTC fed laying hens i.e. (number of positive samples/number of samples tested)×100.</p

The PCR products from the tissues of laying hens fed diets containing control corn (CC) and phytase transgenic corn (PTC) for 16 weeks.

<p>(a) The endogenous gene (396 bp) of poultry for <i>ov</i> (left), (b) The exogenous gene (678 bp) of PTC for <i>phy</i>A2 (right). Arrows indicate the expected length of PCR products. M = marker; Lanes 1∼6 = blood, heart, liver, spleen, kidney, breast muscle of CC-fed laying hens (control); lanes 7∼12 = blood, heart, liver, spleen, kidney, breast muscle of PTC-fed laying hens; − = negative control (no DNA).</p

The PCR products from the gastrointestinal contents of laying hens fed diets containing control corn (CC) and phytase transgenic corn (PTC) for 16 weeks.

<p>(a) The endogenous gene (226 bp) of corn for <i>ivr</i> (left), (b) The exogenous gene (678 bp) of PTC for <i>phy</i>A2 (right). Arrows indicate the expected length of PCR products. M = marker; lanes 1 to 8 CC-fed laying hens (control); lanes 9 to 16 = PTC-fed laying hens; + = positive (GM corn), and − = negative control (no DNA). C = crop contents, G = gizzard contents, D = duodenal contents, J = jejunum contents, I = ileum contents, CE = cecal contents, and R = rectal contents.</p

Detection of the transgenic <i>phy</i>A2 gene in corn, diet, digesta, tissues, blood, and eggs of laying hens fed diets containing control corn (CC) and phytase transgenic corn (PTC) for 16 weeks¹.

1<p>Number of samples that tested positive for the <i>phy</i>A2 gene out of 8 samples analyzed. One sample was tested per laying hen (n = 8 laying hens per treatment).</p>2<p>Percentage of samples positive for the <i>phy</i>A2 gene taken from PTC fed laying hens i.e. (number of positive samples/number of samples tested)×100.</p

Composition of the diets (as-fed basis).

1<p>CC = control corn; PTC = phytase transgenic corn.</p>2<p>Provided per kilogram of diet: vitamin A, 12,200 IU; vitamin D3, 4,125 IU; vitamin E, 30 IU; vitamin K, 4.5 mg; thiamine, 1 mg; riboflavin, 8.5 mg; calcium pantothenate, 50 mg; niacin, 32.5 mg; pyridoxine, 8 mg; biotin, 2 mg; folic acid, 5 mg; vitamin B12, 5 mg; manganese, 80 mg; iodine, 1 mg; iron, 60 mg; copper, 8 mg; zinc, 80 mg; selenium, 0.3 mg.</p>3<p>Calculated values.</p>4<p>Determined values.</p

Western blot identification of <i>phy</i>A2 protein in gastrointestinal digesta of laying hens fed diet phytase transgenic corn for 16 weeks.

<p>“+” Positive control (GM corn); lane 1, crop contents; lane 2, gizzard contents; lane 3, duodenal contents; lane 4, jejunum contents; lane 5, ileum contents; lane 6, cecal contents; lane 7, rectal contents.</p

Performance and egg quality of laying hens fed diets containing control corn (CC) and phytase transgenic corn (PTC) for 16 weeks¹.

1<p>Data are expressed as mean ± SD (<i>n</i> = 8).</p>a,b<p>Means within a row with no common superscripts differ significantly (<i>P</i><0.05).</p

Fragmentation of DNA from control corn (CC) and phytase transgenic corn (PTC).

<p>(a) The endogenous gene (226 bp) of corn for <i>ivr</i> (left), (b) The exogenous gene (678 bp) of PTC for <i>phy</i>A2 (right). Arrows indicate the expected length of PCR products. M = marker; Lanes 1 = CC; lanes 2 = PTC; Lanes 3 = CC diet; lanes 4 = PTC diet; − = negative control (no DNA).</p

Comparison of amino acid sequences of HA and NA of H9N2 viruses tested in this study

a<p>RBS, receptor binding site.</p>b<p>Amino acid sequence of residues 324–330 (numbering according to H3 HA) at the cleavage site between HA1 and HA2; aa, amino acid.</p