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Home / Issues / № 2, 2015

Biological sciences

DEVELOPMENT OF PRIMERS AND PROBE FOR HORSEFLESH PCR IDENTIFICATION IN MEAT PRODUCTS.
Aubekerova L. S., Sarbakanova SH. T., Minayeff M. Yu.

Introduction Ensuring reliable food security of the country, certainly, is today one of the major, priority national objectives. Health and welfare of the nation, economic and political independence directly depends on its successful decision. Lack of quality control and safety of meat products can lead to use of raw materials of lower grade, falsification of one type of meat by another [1, 2, 3].

Materials and methods. To achieve the objectives were used molecular genetic techniques. A detailed analysis of the mitochondrial DNA of the studied biological objects and search for their nucleotide sequences were carried out on the NCBI. Analysis of the variability of the selected nucleotide sequences and searching conserved regions required for primer selection was carried out using computer programs CLC Sequence Viewer 2 and Primer Express. The specificity of the designed primers theoretically studied using an interactive system BLAST on-line.

The results of research. To find sequences of genomes of animals necessary to us in the section "Nucleotide" enter the name of a genome of an animal necessary to us, we enter on NCBI base "Equus caballus cytb".

In the results offered by the program to us, we choose nucleotide sequences necessary to us, the consisting about 1000, and in the following link we see full sequence of "Equus caballus". Further, these sequencings write out for carrying out alignment with other sequenced sequences and place in the BLAST program. Choose some sequencings of a gene of cytochrome in a horse with the biggest divergence in nucleotide sequence from the given search results, in a genome of a horse it made 3%, that is 97% are an intraspecific specificity in different the sequencing cytochrome gene options in a horse.

For selection of primers enter the selected sequences into the CLC Sequence Viewer program. The program selects some options of primers from which in the Oligo Analyzer program, according to the necessary characteristics, one direct (forward) primer is selected. In addition, in this program selection of the return (reverse) primer to the forward begins the way stated above.

The primers answering to all above requirements and the most optimum for identification of specific accessory of meat of a horseflesh and meat ingredients in composition of foodstuff were previously chosen from the options given by the program.

For a horse (Equus caballus) the chosen primer consisted of the following nucleotide sequences:

Direct primer: 5-GACTCCTCCTCCTGATCTTGCT-3';

Return primer: 5-GTAGGATGGCGTAGGCAAACAG-3';

Probe: 5-CCAGCTAACCCTCTCAGCAC-3'.

Conclusions In such way, in result of researches specific nucleotide sequences of the studied DNA of a horse are defined, the perspective matrix in the form of a gene cytochrome B is chosen in and the design the specific the oligonucleotide of primers for RT PCR of identification of DNA of a horse (Equus caballus) is carried out.



References:
1. Gordeev, A.V. Food security of Russia / A. V. Gordeev//Innovative technologies and equipment for the food industry (development priorities): materials III international scientific and practical conference. – Voronezh: Publisher of VGTA, 2009. – Page 584.

2. Kuzmicheva, M. B. Food security – an important factor of economic growth of the state / M. B. Kuzmicheva//the Meat industry. –2010. – No. 1. – Page 4-7.

3. Fei S., Okayama T.,Yamanoue M., Nishikawa I., Mannen H., Tsuji S. Species identification of meats and meat products by PCR // Anim. Sc. Technol., 1996. V.61, № 10. P. 900-905.



Bibliographic reference

Aubekerova L. S., Sarbakanova SH. T., Minayeff M. Yu. DEVELOPMENT OF PRIMERS AND PROBE FOR HORSEFLESH PCR IDENTIFICATION IN MEAT PRODUCTS. . International Journal Of Applied And Fundamental Research. – 2015. – № 2 –
URL: www.science-sd.com/461-24921 (29.03.2024).