The sweet potato ADP-glucose pyrophosphorylase gene (ibAGP1) promoter confers high-level expression of the GUS reporter gene in the potato tuber.
Molecular farming refers to the process of creating bioengineered plants with the capability of producing potentially valuable products, such as drugs, vaccines, and chemicals. We have investigated the potential of the sweet potato ADP-glucose pyrophosphorylase gene (ibAGP1) promoter and its transit peptide (TP) as an expression system for the mass production of foreign proteins in potato. The ibAGP1 promoter and its TP sequence were transformed into potato along with beta-glucuronidase (GUS) as a reporter gene, and GUS activity was subsequently analyzed in the transgenic potato plants. In tuber tissues, GUS activity in transgenic plants carrying only the ibAGP1 promoter (ibAGP1::GUS) increased up to 15.6-fold compared with that of transgenic plants carrying only the CaMV35S promoter (CaMV35S::GUS). GUS activity in transgenic plants was further enhanced by the addition of the sweetpotato TP to the recombinant vector (ibAGP1::TP::GUS), with tuber tissues showing a 26-fold increase in activity compared with that in the CaMV35S::GUS-transgenic lines. In leaf tissues, the levels of GUS activity found in ibAGP1::GUS-transgenic lines were similar to those in CaMV35S::GUS-lines, but they were significantly enhanced in ibAGP1::TP::GUS-lines. GUS activity gradually increased with increasing tuber diameter in ibAGP1::GUS-transgenic plants, reaching a maximum level when the tuber was 35 mm in diameter. In contrast, extremely elevated levels of GUS activity - up to about 10-fold higher than that found in CaMV35S::GUS-lines - were found in ibAGP1::TP::GUS-transgenic lines at a much earlier stage of tuber development (diameter 4 mm), and these higher levels were maintained throughout the entire tuber developmental stage. These results suggest that the sweetpotato ibAGP1 promoter and its TP are a potentially strong foreign gene expression system that can be used for molecular farming in potato plants.
Kim
Tae-Won
TW
College of Life Science & Natural Resources, JinJu National University, JinJu 660-758, Republic of Korea.
Goo
Young-Min
YM
Lee
Cheol-Ho
CH
Lee
Byung-Hyun
BH
Bae
Jung-Myung
JM
Lee
Shin-Woo
SW
D016960:Agrobacterium tumefaciens / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000235:genetics*; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D051599:Glucose-1-Phosphate Adenylyltransferase / Q000235:genetics* / Q000502:physiology; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics*; D027723:Ipomoea batatas / Q000201:enzymology / Q000235:genetics*; D018515:Plant Leaves / Q000201:enzymology; D010940:Plant Proteins / Q000235:genetics* / Q000502:physiology; D035281:Plant Tubers / Q000201:enzymology* / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D018749:RNA, Plant / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D011198:Solanum tuberosum / Q000201:enzymology* / Q000254:growth & development; D013045:Species Specificity; D013395:Sucrose / Q000494:pharmacology; D014170:Transformation, Genetic
D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; D011993:Recombinant Fusion Proteins; D013395:Sucrose; D051599:Glucose-1-Phosphate Adenylyltransferase; D005966:Glucuronidase