Estimating number of transgene copies in transgenic rapeseed by real-time PCR assay with HMG I/Y as an endogenous reference gene

Plant Molecular Biology Reporter

Volumn 22, Issue 3, 2004, Pages 289-300

  Weng, Haibo ^a,b   Pan, Aihu ^c   Yang, Litao ^a   Zhang, Chengmei ^c   Liu, Zhili ^a   Zhang, Dabing ^b,c  

^a NANJING UNIVERSITY   (China)

^b SHANGHAI JIAO TONG UNIVERSITY   (China)

^c INSTITUTE OF EDIBLE FUNGI   (China)

Author keywords

Brassica napus; HMG I Y; Real time PCR; Transgene copy number

Indexed keywords

BRASSICA NAPUS; SINAPIS ARVENSIS;

EID: 13444311862     PISSN: 07359640     EISSN: 07359640     Source Type: Journal    
DOI: 10.1007/BF02773139     Document Type: Article

References (22)

1. Callaway AS, Abranches R, Scroggs J, Allen GC, and Thompson WF (2002) Highthroughput transgene copy number estimation by competitive PCR. Plant Mol Biol Rep 20: 265-277.
2. Celi FS, Cohen MM, Antonarakis SE, Wertheimer E, Roth J, and Shuldiner AR (1994) Determination of gene dosage by a quantitative adaptation of the polymerase chain reaction (gd-PCR): rapid detection of deletions and duplications of gene sequences. Genomics 21: 304-310.
3. Chengmei Z, Bingjun Q, Yun X, Yahong H, Aihu P, Wanqi L, and Dabing Z (2003) Transferring an antisense acyl-acyl carrier protein (ACP) DNA fragment into Brassica napus by high efficient Agrobacterium tumefaciens genetic transformation system. Acta Agriculturae Shanghai 19: 5-8.
4. De Preter K, Speleman F, Combaret V, Lunec J, Laureys G, Eussen BH, Francotte N, Board J, Pearson AD, De Paepe A, Van Roy N, and Vandesompele J (2002) Quantification of MYCN, DDX1, and NAG gene copy number in neuroblastoma using a realtime quantitative PCR assay. Mod Pathol 15: 159-166.
5. Flavell RB (1994) Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc Natl Acad Sci USA 91: 3490-3496.
6. Ginzinger DG (2002) Gene quantification using real-time quantitative PCR: an emerging technology hits the mainstream. Exp Hematol 30: 503-512.
7. Huang Y, Liang W, Pan A, Zhou Z, Huang C, Chen J, and Zhang D (2003) Production of FaeG, the major subunit of K88 fimbriae, in transgenic tobacco plants and its immunogenicity in mice. Infect Immun 71: 5436-5439.
8. Ingham DJ, Beer S, Money S, and Hansen G (2001) Quantitative real-time PCR assay for determining transgene copy number in transformed plants. Biotechniques 31: 136-140.
9. Iyer LM, Kumpatla SP, Chandrasekharan MB, and Hall TC (2000) Transgene silencing in monocots. Plant Mol Biol 43: 323-346.
10. Jiayu D, Junwei J, Litao Y, Weng H, Chengmei Z, Wenxuan L, and Dabing Z (2004) Validation of a rice specific gene, sucrose phosphate synthase, used as the endogenous reference gene for qualitative and real-time quantitative PCR detection of transgenes. J Agric Food Chem 52: 3372-3377.
11. Khachatourians GG, McHughen A, Scorza R, Nip W, and Hui Y (2002) Transgenic plants and crops. Marcel Dekker, New York.
12. Kooter JM, Matzke TA, and Meyer P (1999) Listening to the silent genes: transgene silencing, gene regulation, and pathogen control. Trends Plant Sci 4: 340-347.
13. -ΔΔCT method. Methods 25: 402-408.
14. Luo ZY, Zhou G, and Chen XH (2001) Isolation of high-quality genomic DNA from plants. Hunan Yi Ke Da Xue Xue Bao 26: 178-180.
15. Masek T, Smykal P, Janotova II, Honys D, Capkova VV, and Pechan PM (2002) Isolation of a Brassica napus L. cDNA encoding a putative high-mobility-group HMG I/Y protein. Plant Sci 159: 197-204.
16. Mason G, Provero P, Vaira AM, and Accotto GP (2002) Estimating the number of integrations in transformed plants by quantitative real-time PCR. BMC Biotechnol 2: 20.
17. Murray MG and Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8: 4321-4325.
18. Sambrook J, Fritsch EF, and Maniatis T (1989) Molecular cloning: a laboratory manual. 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
19. TM-derived transgenic maize. Plant Cell Rep 20: 948-954.
20. Vaucheret H, Beclin C, Elmayan T, Feuerbach F, Godon C, Morel JB, Mourrain P, Palauqui JC, and Vernhettes S (1998) Transgene-induced gene silencing in plant. Plant J 16: 651-659.
21. Weng H, Yang L, Liu Z, Ding J, Pan A, and Zhang D (2004) A novel reference gene, high-mobility-group protein I/Y, can be used in qualitative and real-time quantitative PCR detection of transgenic rapeseed cultivars. J AOAC Int. Forthcoming.
22. Zhang Y, Zhang D, Li W, Chen J, Peng Y, and Cao W (2003) A novel real-time quantitative PCR method using attached universal template probe. Nucleic Acids Res 31: e123.