Qiu Youwen

Establishment of TaqMan Real-time Quantitative PCR Assay for Foreign Gene Copy Numbers in Transgenic Soybean

Abstract TaqMan quantitative PCR technique was used to detect the copies of exogenous CaMV35S flanks sequence in transgenic soybean. With soybean lectin as the endogenous reference gene, and gene complex DNA in non-GMO soybeans as the endogenous reference standard, the gradient dilution method was used to separately calculate C t value of endogenous reference gene and plasmid DNA and correlation standard curve equation of logarithm of copies, and then to calculate the copies of samples through substituting thus-obtained C t into the standard curve equation. The standard curve equation of endogenous reference gene was y =−3.422x+35.201, R 2 =0.998; the standard curve equation of exogenous gene was y =−3.495x+35.303, R 2 =0.999. The sample copies was got by putting C t value into the standard curve equation, and it was the ratio of exogenous gene and reference gene. We found that CaMV35S gene in transgenic soy was single copy.

Study on Lysine and Methionine Content Promotion of Soybean Meal by Probiotic Fermentation Process

Abstract Soybean meal (SBM) is commonly used for livestock feeds, but its application in diets for livestock is limited due to some antinutritional factors. The contents of methionine and lysine of soybean meal were promoted by Bacillus natto and Leuconostoc mesenteroides fermentation, benefial for the livestock feeds. It was crude protein (CP) 56.8%, methionine 43.56 mg • g −1 , and lysine 74.87 mg • g −1 , cows fed a diet with FSBM milk yield raised 14.2%, the change in the milk protein, the lactose and the dry matter content had also obvious increase. This convenient technique offers helpful exploration for industrialization of soybean meal fermentation.

SYBR® Green qPCR Screening Methods for Detection of Anti-herbicide Genes in Genetically Modified Processed Products

The use of genetically modified organisms (GMOs) as food products becomes more and more widespread. The European Union has implemented a set of very strict procedures for the approval to grow, import and/or utilize GMOs as food or food ingredients. Thus, analytical methods for detection of GMOs are necessary in order to verify compliance with labelling requirements. There are few effective screening methods for processed GM (genetically modified) products. Three anti-herbicide genes (CP4-EPSPS, BAR and PAT) are common exogenous genes used in commercialized transgenic soybean, maize and rice. In the present study, a new SYBR® Green qPCR screening method was developed to simultaneously detect the three exogenous anti-herbicide genes and one endogenous gene in a run. We tested seven samples of representative processed products (soya lecithin, soya protein powder, chocolate beverage, infant rice cereal, maize protein powder, maize starch, and maize jam) using the developed method, and amplicons of endogenous gene and transgenic fragments were obtained from all the processed products, and the sensitivity was 0.1%. These results indicated that SYBR® Green qPCR screening method was appropriate for qualitative detection of transgenic soybean, maize and rice in processed products.

Establishment of PCR-ELISA for Detecting Glyphosate Resistant Transgenic Soybean

Abstract A PCR-ELISA method for detecting the glyphosate resistant transgenic soybean was established and optimized. The results showed that the key parameters of PCR-ELISA were as follows: the concentration of digoxin tag probe was 0.5 µmol · L −1 , the time of hybridization reaction was 15 min and the chromogenic reaction should last for 30 min. The sensitivity and the repeatability of our PCR-ELISA method were evaluated, and the results showed that it could be detected when the concentration of DNA template from transgenic soybean samples was 0.01% or higher, and the coefficient of variation of this method was less than 5% in our research condition. These results suggested that PCR-ELISA method establishment in this study had good repeatability and high precision for detecting the transgenic soybean samples.

Screen of Bovine Mammary Gland Epithelial Cell Specifcity Promotor

Three lactoproteins (α-S1-casein, β-lactoglobulin, and β-casein) promotors were cloned, sequenced and compared relative luciferase expression. The results showed that the promotor activity of bovine α-S1-casein gene was the best, and would be used to produce pharmaceutically and medically important proteins in the mammary gland of transgenic animals and also for the construction of an inducible eukaryotic expression vector.