EUROMEDICA  Hanover 6-7  Juni 2008	Advanced methods of diagnosis, treatment and prophylactics
	European Academy of Natural Sciences, Hanover European Scientific Society, Hanover Russian Academy of Natural Sciences, Moscow

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The process of gene expression involves several steps: transcription, RNA processing, translation and assembly of protein complexes. In each of these steps it is possible to take control over gene expression, both on the part of the genome, and from the abiogenous factors. Formation of lactic acid bacteria is of practical importance, as a part of the antimicrobial mechanism. Study of mechanisms of acid production is actual for biochemical use, as well as for genetic methods, which give us the opportunity to manage the processes of acid production. The aim of our research is to study the mechanisms of gene expression responsible for the production of lactic acid.

As the object of studies strains of Lactobacillus casei were used, which characterize by common for this type of morphological, cultural and biochemical properties. Bacterial strains are isolated from different sources in different regions of Kazakhstan, typing by sequencing as Lactobacillus casei. In this paper we used strains of Lactobacillus casei possesing by different levels of acid production

To determine the expression of lactate dehydrogenase genes it was carried out two step RT Real-Time PCR. As intercalating agent it was used SybrGreenI. To determine the initial number of cDNA sample it was used the algorithm describing the behavior of the kinetic curve. As a criterion of normal distribution it was using the Shapiro-Vilkonsona test, using the expression of “house keeping” genes.

The graph for determining the melting curves of the gene expression of lactate dehydrogenase had a peak with the melting point, indicating that only the accumulation of specific products.

The maximum value acid activity of Lactobacillus casei correspond to the maximum level of lactate dehydrogenase gene. The coefficient of correlation is 0.25, indicating a linear correlation of two traits.

Determined the levels of lactate dehydrogenase mRNA by real-time PCR. It was found, the level of lactate dehydrogenase mRNA varies slightly. Decrease in the acid activity was characterized by low levels of mRNA compared with control: lower by 63 - 76%, while increasing the rate of acid activity was characterized by high levels of mRNA: 11 - 17% of the level of the benchmarks. The data obtained are preliminary and do not give the full specifications of acid production depending on the lactate dehydrogenase gene expression.