Real-Time PCR Method Using Capturing Oligo-Immobilized PCR Tubes to Determine the Specific Gene for Soybean and Genetically Modified Soybean in Food Matrices

被引：3

作者：

Harikai, Naoki ^{[1
]}

Saito, Shin ^{[2
]}

Abe, Midori ^{[2
]}

Kondo, Kazunari ^{[3
]}

Kitta, Kazumi ^{[4
]}

Akiyama, Hiroshi ^{[3
]}

Teshima, Reiko ^{[3
]}

Kinoshita, Kenji ^{[1
]}

机构：

[1] Mukogawa Womens Univ, Sch Pharmaceut Sci, Nishinomiya, Hyogo 6638179, Japan

[2] Sumitoma Bakelite Co, Nishi Ku, Kobe, Hyogo 6512241, Japan

[3] Natl Inst Hlth Sci, Setagaya Ku, Tokyo 1588501, Japan

[4] Natl Agr & Food Res Org, Natl Food Res Inst, Tsukuba, Ibaraki 3058642, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2008年 / 72卷 / 11期

关键词：

DNA extraction; genetically modified organism; hybridization; polymerase chain reaction; single-tube method;

D O I：

10.1271/bbb.80433

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A new real-time PCR method using capturing oligo-immobilized PCR tubes is described. This method was used to detect specific genes for soybean and genetically modified (GM) soybean in food matrices. In a standard reaction using soybean genomic DNA and a capturing oligo for the lectin gene (Le1) immobilized on the tube, we examined the effects of such hybridization conditions as the location, length, and amount of the capturing oligo, and the incubation time and temperature. Under optimized conditions, the copy number of Le1 was determined in a concentration-dependent manner from soybean genomic DNA and soybean lysate (DNA 101000 ng, r = 0.99; lysate 1-100%, r = 0.99). The copy number of a Roundup Ready soybean (RRS) gene was also successfully detected in a concentration-dependent manner (1-100%, r = 0.99) from GM soybean lysate, using PCR tubes with an immobilized capturing oligo for the transgene. Our data indicate that this is a rapid and simple method to determine specific genes for soybean and GM soybean in food matrices.

引用

页码：2953 / 2958

页数：6

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