Fig. 4
From: Expression of recombinant human Apolipoprotein A-IMilano in Nicotiana tabacum
A PCR confirmation of the presence of the Apo A-IMilano gene and 3xFlag (934 bp) in stable transgenic N. tabacum leaves. Lane M is 2000 bp DNA ladder. Lane NC is the negative control. Lanes 1 to 17 are the PCR products of antibiotic-resistant N. tabacum plants, p478 represents the batch number of the stable transgenic N. tabacum plants, and numbers 1 to 17 in the PCR test results correspond to p478-9, p478-35, p478-39, p478-53, p478-63, p478-64, p478-67, p478-69, p478-73, p478-7, p478-13, p478-14, p478-21, p478-32, p478-51, p478-56, and p478-74 plants, respectively, These plants are the number of 17 Apo A-IMilano DNA-positive individual plants selected from 86 transgenic N. tabacum plants in the same batch p478. Lane PC was a positive control. B RT-qPCR analysis of stably transgenic N. tabacum leaves from 13 plants with positive mRNA expression were screened from 17 plants with positive DNA detection in the PCR test results. These were p478-9, p478-35, p478-39, p478-53, p478-63, p478-7, p478-13, p478-14, p478-21, p478-32, p478-51, p478-56, p478-74 plants. C Western blotting of transformants. Lane M is protein molecular weight marker. Lane NC presents wild N. tabacum. Lanes 2 to 9 present recombinant fusion expression of Flag–Apo A-IMilano in stable transgenic N. tabacum with a molecular weight of 30 KDa. Lane PC presents the standard protein Apo A-I with a molecular weight of 28 KDa. Numbers 2 to 9 in the western blot results correspond to numbers of p478-51, p478-32, p478-53, p478-7, p478-9, p478-63, p478-35, p478-56 plants, respectively. These are 8 plants with positive expression of Apo A-IMilano protein selected from 13 plants with positive mRNA expression of N. tabacum