Abstract
The development of molecular genetic and bioinformatic systems for identifying the species of milk and the raw material composition of dairy products is of great scientific and practical importance with the purpose of introducing developments in the system for controlling the turnover of falsified products. The aim of the research is to develop a method of PCR-RFLP analysis for species identification of milk and dairy products from agricultural ruminant animals by the κ-casein gene (CSN3) with the possibility of qualitative and relative quantitative assessment of species-specific DNA of the tested biomaterial. The objects of research were samples of raw milk and milk powder, pasteurized cream, and hard and semi-hard cheeses. The developed method of species identification of milk and dairy products includes sample preparation of the studied samples, nucleic acid extraction, combined PCR-RFLP technique, detection of obtained results by the method of horizontal electrophoresis in agarose gel and their analysis, including using the developed mathematical algorithms and software. The synergistic effect established in combined operation of 2 restriction enzymes ensured their application in a mix with increased performance in an ergonomic way in the context of DNA authentication of cow, goat, and sheep milk and dairy products based on them. The specificity and sensitivity of the proposed method is potentially suitable for implementing the development of a system to control the turnover of falsified and counterfeit goods.
Highlights
Regarding the DNA authentication of milk and dairy products, the first step is the identification of the type of milk and the raw material composition of dairy products, sourced from the main set of dairy animal species, which together account for 99% of world milk production
Additional authentication approaches based on the length polymorphism of restriction fragments of amplified DNA (PCR-RFLP) have found their application along with methods of hybridization, sequence, and biosensor analysis of the corresponding nucleotide sequences of mitochondrial and nuclear genes
Species identification in milk and raw material composition of dairy products was performed based on the developed PCR-RFLP technique for the analyzed locus of the κ-casein gene of agricultural ruminant animals
Summary
In the modern food system one of the most important aspects is the problem of identification of food raw Regarding the DNA authentication of milk and dairy products, the first step is the identification of the type of milk and the raw material composition of dairy products, sourced from the main set of dairy animal species, which together account for 99% of world milk production. The technological accessibility and simplicity of the PCR-RFLP, as well as its analytical characteristics (sensitivity, specificity, and reproducibility), ensure the correct interpretation of the results This informative authentication method is diagnostically significant and popular, despite the relatively long procedure for endonuclease cleavage of the synthesized PCR product (Hashim and Al-Shuhaib, 2019). Among the many identification molecular markers of nuclear DNA, the genes of milk proteins linked to economically valuable traits of animals deserve attention (Tyulkin, 2018). It is the κ-casein gene (CSN3), whose intra- and interspecies genetic polymorphism is the most studied and is widely represented in the form of deposited nucleotide sequences in bioinformatics databases
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