PMID- 16076097
OWN - NLM
STAT- MEDLINE
DCOM- 20050919
LR  - 20201209
IS  - 0021-8561 (Print)
IS  - 0021-8561 (Linking)
VI  - 53
IP  - 16
DP  - 2005 Aug 10
TI  - Identification and quantification of three genetically modified insect resistant 
      cotton lines using conventional and TaqMan real-time polymerase chain reaction 
      methods.
PG  - 6222-9
AB  - As the genetically modified organisms (GMOs) labeling policies are issued in many 
      countries, qualitative and quantitative polymerase chain reaction (PCR) 
      techniques are increasingly used for the detection of genetically modified (GM) 
      crops in foods. Qualitative PCR and TaqMan real-time quantitative PCR methods to 
      detect and identify three varieties of insect resistant cotton, i.e., Mon531 
      cotton (Monsanto Co.) and GK19 and SGK321 cottons (Chinese Academy of 
      Agricultural Sciences), which were approved for commercialization in China, were 
      developed in this paper. Primer pairs specific to inserted DNAs, such as Cowpea 
      trypsin inhibitor (CpTI) gene of SGK321 cotton and the specific junction DNA 
      sequences containing partial Cry1A(c) gene and NOS terminator of Mon531, GK19, 
      and SGK321 cotton varieties were designed to conduct the identified PCR assays. 
      In conventional specific identified PCR assays, the limit of detection (LOD) was 
      0.05% for Mon531, GK19, or SGK321 in 100 ng of cotton genomic DNA for one 
      reaction. Also, the multiplex PCR method for screening the three GM cottons was 
      also established, which could save time and cost in practical detection. 
      Furthermore, a real-time quantitative PCR assay based on TaqMan chemistry for 
      detection of insect resistant gene, Cry1A(c), was developed. This assay also 
      featured the use of a standard plasmid as a reference molecule, which contained 
      both a specific region of the transgene Cry1A(c) and an endogenous stearoyl-acyl 
      carrier protein desaturase (Sad1) gene of the cotton. In quantitative PCR assay, 
      the quantification range was from 0.01 to 100% in 100 ng of the genome DNA 
      template, and in the detection of 1.0, 3.0, and 5.0% levels of three insect 
      resistant cotton lines, respectively, all of the relative standard deviations 
      (RSDs) were less than 8.2% except for the GM cotton samples with 1.0% Mon531 or 
      GK19, which meant that our real-time PCR assays involving the use of reference 
      molecule were reliable and practical for GM insect resistant cottons 
      quantification. All of these results indicated that our established conventional 
      and TaqMan real-time PCR assays were applicable to detect the three insect 
      resistant cottons qualitatively and quantitatively.
FAU - Yang, Litao
AU  - Yang L
AD  - School of Life Science and Biotechnology, Shanghai Jiao Tong University, 800 
      Dongchuan Road, Shanghai 200240, People's Republic of China.
FAU - Pan, Aihu
AU  - Pan A
FAU - Zhang, Kewei
AU  - Zhang K
FAU - Guo, Jinchao
AU  - Guo J
FAU - Yin, Changsong
AU  - Yin C
FAU - Chen, Jianxiu
AU  - Chen J
FAU - Huang, Cheng
AU  - Huang C
FAU - Zhang, Dabing
AU  - Zhang D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Agric Food Chem
JT  - Journal of agricultural and food chemistry
JID - 0374755
RN  - 0 (Bacillus thuringiensis Toxins)
RN  - 0 (Bacterial Proteins)
RN  - 0 (Bacterial Toxins)
RN  - 0 (DNA, Plant)
RN  - 0 (Endotoxins)
RN  - 0 (Hemolysin Proteins)
RN  - 0 (Trypsin Inhibitors)
RN  - 0 (insecticidal crystal protein, Bacillus Thuringiensis)
SB  - IM
MH  - Animals
MH  - Bacillus thuringiensis Toxins
MH  - Bacterial Proteins/genetics
MH  - Bacterial Toxins/genetics
MH  - DNA, Plant/*analysis
MH  - Endotoxins/genetics
MH  - Fabaceae/chemistry/genetics
MH  - Gossypium/*genetics
MH  - Hemolysin Proteins
MH  - Insecta
MH  - Pest Control, Biological
MH  - Plants, Genetically Modified/*genetics
MH  - Polymerase Chain Reaction/*methods
MH  - Trypsin Inhibitors/genetics
EDAT- 2005/08/04 09:00
MHDA- 2005/09/20 09:00
CRDT- 2005/08/04 09:00
PHST- 2005/08/04 09:00 [pubmed]
PHST- 2005/09/20 09:00 [medline]
PHST- 2005/08/04 09:00 [entrez]
AID - 10.1021/jf050095u [doi]
PST - ppublish
SO  - J Agric Food Chem. 2005 Aug 10;53(16):6222-9. doi: 10.1021/jf050095u.