PCR Detection Method for Transgenic Maize KJ1003

doi:10.19586/j.2095-2341.2025.0179

Current Biotechnology ›› 2026, Vol. 16 ›› Issue (4): 843-852.DOI: 10.19586/j.2095-2341.2025.0179

• Special Forum on Detection Technology for Genetically Modified Organisms • Previous Articles Next Articles

PCR Detection Method for Transgenic Maize KJ1003

Fangrui GAO¹(), Ziyan CHEN¹, Tongtong ZHAO¹, Hongye JIANG¹, Hua ZHANG¹, Yichen LIAO¹, Xiaodan JIANG¹, Fangqi FU¹, Yahui LI¹^,², Haoqian WANG¹(), Hong CHEN¹()

^1.Development Center of Science and Technology，Ministry of Agriculture and Rural Affairs，Beijing 100176，China
^2.Institute of Food Safety and Nutriton，Jiangsu Academy of Agricultural Sciences，Nanjing 210014，China

Received:2025-12-15 Accepted:2026-01-26 Online:2026-07-25 Published:2026-09-11
Contact: Haoqian WANG,Hong CHEN

Abstract

Abstract:

The genetically modified maize KJ1003 is a strain developed by Kejidalong （Beijing） Biotechnology Co.， Ltd. It shows insect resistance and herbicide tolerance has obtained a genetically modified organism safety certificate and has a good industrialization prospect. This study established a qualitative PCR detection method for maize KJ1003， through primer design， primer screening， specificity test， sensitivity test， detection limit test， stability test， and laboratory validation， with the specific sequence of KJ1003 as the target. Results showed that a 165 bp specific fragment could be amplified， with a detection sensitivity of 0.05% and a limit of detection of 0.1%， and had a good stability. Verification results of specificity， detection limit， repeatability and reproducibility for this method from 8 laboratories all met the national standard requirements. This study provides a scientific basis and technical support for the detection and safety supervision of genetically modified crops in China.

Key words: genetically modified maize, event KJ1003, anti-insect and herbicide-tolerant, event-specific, gel-based PCR, qualitative detection

CLC Number:

Fangrui GAO, Ziyan CHEN, Tongtong ZHAO, Hongye JIANG, Hua ZHANG, Yichen LIAO, Xiaodan JIANG, Fangqi FU, Yahui LI, Haoqian WANG, Hong CHEN. PCR Detection Method for Transgenic Maize KJ1003[J]. Current Biotechnology, 2026, 16(4): 843-852.

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URL: https://swjsjz.magtechjournal.com/EN/10.19586/j.2095-2341.2025.0179

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Figures/Tables 8

References 27

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转基因混合样品	组成成分(每种转化体质量分数1%)
其他转基因玉米混合样	Bt11、Bt176、MON810、MON863、GA21、NK603、T25、TC1507、MON89034、MON88017、59122、MIR604、3272、MON87460、DAS40278-9、4114、MON87427、5307、DBN9858、ND207、DBN9501、瑞丰125、DBN9936、MON87411、MZIR098、瑞丰8、BFL4-2、nCX-1
转基因大豆混合样	GTS40-3-2、MON89788、A5547-127、A2704-12、356043、305423、CV127、MON87701、MON87708、MON87769、MON87705、FG72、DAS81419-2、DAS-444Ø6-6、MON87751、SHZD3201、CAL16、中黄6106、DBN9004
转基因水稻混合样	TT51-1、KF-6、KMD-1、M12、KF-8、KF-2、G6H1、T1C-19
转基因油菜混合样	MS1、MS8、RF1、RF2、RF3、T45、Oxy-235、Topas19/2、MON88302、73496
转基因棉花混合样	MON1445、MON531、MON15985、LLCOTTON25、MON88913、GHB614、COT102
非转基因玉米混合样	多种非转基因玉米等量混合制成

转基因混合样品	组成成分(每种转化体质量分数1%)
其他转基因玉米混合样	Bt11、Bt176、MON810、MON863、GA21、NK603、T25、TC1507、MON89034、MON88017、59122、MIR604、3272、MON87460、DAS40278-9、4114、MON87427、5307、DBN9858、ND207、DBN9501、瑞丰125、DBN9936、MON87411、MZIR098、瑞丰8、BFL4-2、nCX-1
转基因大豆混合样	GTS40-3-2、MON89788、A5547-127、A2704-12、356043、305423、CV127、MON87701、MON87708、MON87769、MON87705、FG72、DAS81419-2、DAS-444Ø6-6、MON87751、SHZD3201、CAL16、中黄6106、DBN9004
转基因水稻混合样	TT51-1、KF-6、KMD-1、M12、KF-8、KF-2、G6H1、T1C-19
转基因油菜混合样	MS1、MS8、RF1、RF2、RF3、T45、Oxy-235、Topas19/2、MON88302、73496
转基因棉花混合样	MON1445、MON531、MON15985、LLCOTTON25、MON88913、GHB614、COT102
非转基因玉米混合样	多种非转基因玉米等量混合制成

编号	引物名称及序列（5'→3'）		产物大小/bp	来源
组合1	LB-F1： 5'-CGCGTGAATACGTAGCTGTG-3'	LB-R1： 5'-GAGCGCGTAAGTCTTGGTCT-3'	229	本研究设计
组合2	LB-F2： 5'-GAGCCACACCCAAGTTCCTAA-3'	LB-R2： 5'-TGGCAAATGGGGGATGCTTT-3'	235	本研究设计
组合3	LB-F3： 5'-TGTGTCCCAAGTAAACCTCAGT-3'	LB-R3： 5'-CCAAGATGGAGGGAAGGAGC-3'	253	本研究设计
组合4	LB-F4： 5'-GTAACAGAAAACACCACGGTCC-3'	LB-R4： 5'-ATGAGGGCGACCAGACCTTA-3'	165	本研究设计
组合5	LB-F1： 5'-CGCGTGAATACGTAGCTGTG-3'	LB-R4： 5'-GTAACAGAAAACACCACGGTCC-3'	302	本研究设计
组合6	LB-F2： 5'-GAGCCACACCCAAGTTCCTAA-3'	LB-R1： 5'-GAGCGCGTAAGTCTTGGTCT-3'	131	本研究设计
组合7	LB-F3： 5'-TGTGTCCCAAGTAAACCTCAGT-3'	LB-R2： 5'-TGGCAAATGGGGGATGCTTT-3'	306	本研究设计
组合8	LB-F4： 5'-GTAACAGAAAACACCACGGTCC-3'	LB-R1： 5'-GAGCGCGTAAGTCTTGGTCT-3'	92	本研究设计
组合9	RB-F1： 5'-TTTCTCTTCCCCAACCGTGC-3'	RB-R1： 5'-AGCGCGCAAACTAGGATAAA-3'	143	本研究设计
组合10	RB-F2： 5'-GTGTTCCAGCTGACCGTGT-3'	RB-R2： 5'-CGCGGTGTCATCTATGTTACT-3'	145	本研究设计
组合11	RB-F3： 5'-CGCAGTGCGTTTGAGGC-3'	RB-R1： 5'-AGCGCGCAAACTAGGATAAA-3'	265	本研究设计
组合12	RB-F4： 5'-CATGGCCCTGACCGTAGC-3'	RB-R4： 5'-AAATATAGCGCGCAAACTAGGAT-3'	232	本研究设计
组合13	RB-F2： 5'-GTGTTCCAGCTGACCGTGT-3'	RB-R1： 5'-AGCGCGCAAACTAGGATAAA-3'	173	本研究设计
组合14	RB-F3： 5'-CGCAGTGCGTTTGAGGC-3'	RB-R4： 5'-AAATATAGCGCGCAAACTAGGAT-3'	271	本研究设计
组合15	RB-F4： 5'-CATGGCCCTGACCGTAGC-3'	RB-R2： 5'-CGCGGTGTCATCTATGTTACT-3'	198	本研究设计
组合16	LB-F1： 5'-TCCTCGCTCCCTTTTCTCTT-3'	LB-R1： 5'-TTAGAGTCCCGCAATTATACATT-3'	202	公司提供

编号	引物名称及序列（5'→3'）		产物大小/bp	来源
组合1	LB-F1： 5'-CGCGTGAATACGTAGCTGTG-3'	LB-R1： 5'-GAGCGCGTAAGTCTTGGTCT-3'	229	本研究设计
组合2	LB-F2： 5'-GAGCCACACCCAAGTTCCTAA-3'	LB-R2： 5'-TGGCAAATGGGGGATGCTTT-3'	235	本研究设计
组合3	LB-F3： 5'-TGTGTCCCAAGTAAACCTCAGT-3'	LB-R3： 5'-CCAAGATGGAGGGAAGGAGC-3'	253	本研究设计
组合4	LB-F4： 5'-GTAACAGAAAACACCACGGTCC-3'	LB-R4： 5'-ATGAGGGCGACCAGACCTTA-3'	165	本研究设计
组合5	LB-F1： 5'-CGCGTGAATACGTAGCTGTG-3'	LB-R4： 5'-GTAACAGAAAACACCACGGTCC-3'	302	本研究设计
组合6	LB-F2： 5'-GAGCCACACCCAAGTTCCTAA-3'	LB-R1： 5'-GAGCGCGTAAGTCTTGGTCT-3'	131	本研究设计
组合7	LB-F3： 5'-TGTGTCCCAAGTAAACCTCAGT-3'	LB-R2： 5'-TGGCAAATGGGGGATGCTTT-3'	306	本研究设计
组合8	LB-F4： 5'-GTAACAGAAAACACCACGGTCC-3'	LB-R1： 5'-GAGCGCGTAAGTCTTGGTCT-3'	92	本研究设计
组合9	RB-F1： 5'-TTTCTCTTCCCCAACCGTGC-3'	RB-R1： 5'-AGCGCGCAAACTAGGATAAA-3'	143	本研究设计
组合10	RB-F2： 5'-GTGTTCCAGCTGACCGTGT-3'	RB-R2： 5'-CGCGGTGTCATCTATGTTACT-3'	145	本研究设计
组合11	RB-F3： 5'-CGCAGTGCGTTTGAGGC-3'	RB-R1： 5'-AGCGCGCAAACTAGGATAAA-3'	265	本研究设计
组合12	RB-F4： 5'-CATGGCCCTGACCGTAGC-3'	RB-R4： 5'-AAATATAGCGCGCAAACTAGGAT-3'	232	本研究设计
组合13	RB-F2： 5'-GTGTTCCAGCTGACCGTGT-3'	RB-R1： 5'-AGCGCGCAAACTAGGATAAA-3'	173	本研究设计
组合14	RB-F3： 5'-CGCAGTGCGTTTGAGGC-3'	RB-R4： 5'-AAATATAGCGCGCAAACTAGGAT-3'	271	本研究设计
组合15	RB-F4： 5'-CATGGCCCTGACCGTAGC-3'	RB-R2： 5'-CGCGGTGTCATCTATGTTACT-3'	198	本研究设计
组合16	LB-F1： 5'-TCCTCGCTCCCTTTTCTCTT-3'	LB-R1： 5'-TTAGAGTCCCGCAATTATACATT-3'	202	公司提供

样品名称	8家实验室验证
样品名称	Lab1	Lab2	Lab3	Lab4	Lab5	Lab6	Lab7	Lab8
空白对照	-	-	-	-	-	-	-	-
阴性对照	-	-	-	-	-	-	-	-
阳性对照	+	+	+	+	+	+	+	+
1% KJ1003	+	+	+	+	+	+	+	+
其他转基因玉米混合样	-	-	-	-	-	-	-	-
转基因大豆混合样	-	-	-	-	-	-	-	-
转基因水稻混合样	-	-	-	-	-	-	-	-
转基因油菜混合样	-	-	-	-	-	-	-	-
转基因棉花混合样	-	-	-	-	-	-	-	-
非转基因玉米混合样	-	-	-	-	-	-	-	-
0.1% KJ1003（检出限）	+	+	+	+	+	+	+	+

PCR Detection Method for Transgenic Maize KJ1003

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