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GMO Biosafety Research 2012, Vol.3, No.1, 1-7
http://gmo.sophiapublisher.com
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Figure 1 The insertion fragment of GM cotton 06N-119
amplified by 3′-terminal hiTAIL-PCR
Note: A: The result of first round of
3
′-terminalhiTAIL-PCR
amplification; M: 1kb plus DNA ladder; 1, 2, 3, 4, 5, 6, 7, 8
indicates the amplification product of then 2nd and3rd rounds
of hiTAIL-PCR using four AD primers, AD1-1, AD1-2, AD1-3,
and AD1-4, respectively; B: The result of first round of
3′-terminal LD-PCR amplification; C: The specific amplification
of GM cotton 06N-119; D: The amplification result of the cotton
genomic sequence clos to the 3′-terminal of the insert; M: 1kb plus
DNA ladder; 1: water (Ck); 2: negative control; 3: Zhongmiansuo 41;
4: GK12; 5: Sample
Figure 2 The insertion fragment of GM cotton 06N-119
amplified by 5′-terminal hiTAIL-PCR
Note: A: The result of first round of 5′-terminal hiTAIL-PCR
amplification; M: 1 kb Plus DNA Ladder; 1, 2, 3, 4, 5, 6
indicates the amplification product of the 2nd and3rd rounds of
hiTAIL-PCR; B: The hiTAIL-PCR amplification result of
Non-GM cotton Ji668; M: 1kb plus DNA ladder; 1, 2, 3, 4
indicates the amplification product of 2nd and 3rd rounds of
hiTAIL-PCR; C: The result of 5′-terminal LD-PCR
amplification; M: 1 kb Plus DNA Ladder; 1: Water (CK); 2:
negative control; 3: Zhongmiansuo41; 4: GK12; 5: Sample
of two tandem expression cassettes, namely
Npt II
gene expression cassette and cry1Ac expression
cassette (Figure 3). The structure of the inserting
fragment from 5' to 3' end followed by: (1) ori322/629
bp; (2) oriV/394 bp; (3) E35S promoter/521 bp; (4)
Npt II
/795 bp ; (5) NOS terminator/254 bp; (6) coding
region/789 bp; (7) E35S promoter/521 bp; (8)
cry1Ac
/3537 bp; (9) 7S-UTR/416 bp. The results
further indicated that the promoter for
Npt II
gene was
the enhanced promoter E35S, which was fully