A papaya-specific gene, papain, used as an endogenous reference gene in qualitative and real-time quantitative PCR detection of transgenic papayas

Sensitivity of a real-time PCR method for the detection of transgenes in a mixture of transgenic and non-transgenic seeds of papaya (Carica papayaL.)

“…PCR with forward and reverse primer pairs, viz., papain-A1 and papain-A2 specific for the papaya papain gene, and CP-A1 and CP-A2 for the cp gene (synthesized by Integrated DNA Technologies; http://www.idtdna.com) [22], were used in this study. The papain gene, which is unique to papaya, was used as an internal control, whereas cp gene is found only in transgenic papaya.…”

“…This technique is probably the most appropriate for detecting transgenes in bulked seed lots of papaya. The development of a real-time PCR detection method for assessing transgenic status of papaya using papain and cp genes has been reported [22]. However, the researchers did not report the use of this method on a mixture of GE and non-GE papaya.…”

“…However, the researchers did not report the use of this method on a mixture of GE and non-GE papaya. Thus, our goal was to improve the published method [22] for detection in mixed (GE and non-GE) samples as well as optimize DNA extraction from dried papaya seeds. We envisage our methodology as being helpful to detect adventitious presence of transgenes in mainly non-GE papaya seed lots.…”

Development of Molecular Markers for the Determination of the New Cultivar 'Chunpoong' in Panax ginseng C. A. MEYER Associated with a Major Latex-Like Protein Gene

“…Until now, the real-time PCR method has been successfully used for the detection of celery in food, 18) Phytophthora cryptogea on Gerbera jamesonii, 19) the age of wines, 20) and GM papayas 21) by melting curve analysis. Compared with normal PCR, real-time PCR assay is highly sensitive, specific, fast, and convenient since gel electrophoresis is not needed.…”

A peach (Prunus persica)-specific gene, Lhcb2, used as an endogenous reference gene for qualitative and real-time quantitative PCR to detect fruit products

“…To date, endogenous reference genes of many species have been reported, including the LAT52 gene in the tomato (Yang et al, 2005), SPS in rice (Ding et al, 2004), sad1 in cotton (Xu, Huang, Wang, Zhang, & Luo, 2006), BnACCg8 in rapeseed (Hernandez, Rio, Esteve, Prat, & Pla, 2001), papain in papaya (Xu et al, 2008), the lectin and b-actin genes in the soybean (James, Schmidt, Wall, Green, & Masri, 2003) and the Ivr1, zein, adh1 and hmg genes in maize (Hernandez et al, 2004). Scaravelli, Brohée, Marchelli, and Van Hengel (2008) identified a peanut-specific sequence within the Arah3 allergen gene family; the limit of detection using this sequence is as low as 3 pg DNA.…”