Handbook of Meat Processing

(Greg DeLong) #1

506 Chapter 29


GMO Target sequence Technique Reference
Rapeseed Falcon6/Ac,
HCN10, Liberator6/
Ac, HCN28, HCN92

EPSPS, pat , P - 35S y PEPC RTi - PCR Zeitler et al., 2002

Potato B33 npt II, gbss - as; B33, T - DNA cPCR Hassan - Hauser et al.,
1998
Potato B33 - INV aphIV cPCR Anonymous et al.,
1997
Potato NewLeaf Plus PLRV - rep gene cPCR Hernandez et al.,
2004b
Potato NL Russet
Burbank and NL
Superior

P - 35S and cry 3A cPCR Mel ’ nychuk et al.,
2002

Tomato FlavrSavr ™ npt II, d - P - 35S/
polygalacturonase

cPCR Meyer 1995a, b

Tomato Zeneca T - nos , polygalacturonase cPCR Busch et al., 1999
Maize Bt176, MON810,
Bt11, GA21 and T25)
and soybean Roundup
Ready

Different junctions RTi - PCR fi ve GMO Kuribara et al., 2002

Maize Bt176, MON810,
Bt11, GA21 and
soybean Roundup
Ready

Cp4EPSP/NOS terminator,
35SCamV/BAR, 35ScaMV/
adh, 35ScaMV/hsp70, actin
promoter/CTP

RTi - PCR fi ve GMO Peano et al., 2005

Maize Bt176, Bt11,
GA21, and soybean
GT73

Different junctions RTi - PCR four
GMO

Taverniers et al.,
2005

Maize Bt176, MON810,
Bt11, and soybean
Roundup Ready

Different junctions RTi - PCR four
GMO

Greiner et al., 2005

Maize Roundup ready
and soybean Roundup
ready

CTP/EPSPS RTi - PCR Lerat et al., 2005

Table 29.1. GMO specifi c PCR methods for GMO analysis. cPCR: conventional PCR; RTi-PCR: real-
time PCR; dc-PCR: double competitive PCR; qc-PCR: quantitative competitive PCR. Modifi ed from
Hernandez et al. (2005) (cont.)


ated: a “ nonspecifi c strategy ” independent
of the target sequence (e.g., through fl uores-
cent dyes that have special fl uorescent prop-
erties when bound to double stranded (ds)
DNA), and a “ sequence - specifi c strategy ” by
sequence - specifi c fl uorescent oligonucle-
otide probes. The latter strategy also allows
the simultaneous detection of multiple
targets, using multiple probes labeled with
different reporter dyes (Bernard et al. 1998 ;
Wittwer et al. 2001 ; Dupont et al. 2002 ). This
is of special interest in GMO analysis, since


a wide diversity of transgenic events is intro-
duced in the market.
Several fl uorescent dsDNA - specifi c dyes
have been used in RTi - PCR methods. The fi rst
used was ethidium bromide (Higuchi et al.
1993 ; Le Pecq and Paoletti 1996 ; Wittwer
et al. 1997 ), and subsequently, other interca-
lating dyes have also been introduced, such as
YO - PRO - 1 (Ishiguro et al. 1995 ; Tseng et al.
1997 ) and SYBR Green I (Hernandez et al.
2003c ). They are cost - effective, as it is not
necessary to add a target - specifi c fl uorescent
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