We developed several novel tools to genome wide screen for CNVs and SNPs in single cells. When applied to cleavage stage embryos from young fertile couples we discovered, unexpectedly, an extremely high incidence of chromosomal instability, a hallmark of tumorigenesis (Vanneste et al., Nature Medicine, 2009; Vanneste et al., Hum.Reprod., 2011). Not only mosaicisms for whole chromosome aneuploidies and uniparental disomies but also frequent segmental deletions, duplications and amplifications that were reciprocal in sister blastomeres were detected. Based on the copy number changes that were observed in the blastomeres it was hypothesised that chromosome breakages and fusions occur frequently in cleavage stage human embryos and instigate subsequent breakage-fusion-bridge cycles and that the DNA breaks present in spermatozoa might trigger this CIN. To test these hypotheses, we genotyped both parents as well as 93 blastomeres from 24 IVF embryos and developed a novel SNP-array based algorithm to determine the parental origin of (aberrant) loci in single cells. Paternal as well as maternal alleles were commonly rearranged in the blastomeres indicating that sperm-specific DNA-breaks do not explain the majority of these structural variants. The parent-of-origin analyses together with microarray-guided FISH analyses demonstrate the presence of inv dup del chromosomes as well as more complex rearrangements (Voet et al., Hum.Mutation, 2011). These data provide unequivocal evidence for breakage-fusion-bridge cycles in those embryos and suggest that the human cleavage stage embryo is a major source of chromosomal disorders. The type of rearrangements observed can likely explain the majority of constitutional rearrangements seen in miscarriages as well as live births such as deletions, duplications, inverted deletions duplications, ring chromosomes and mosaicisms of all of those rearrangements. The high frequencies of chromosomal imbalances in cleavage stage embryos make it likely that a substantial number of chromosomal rearrangements originate post-zygoticaly.

Purpose: The aim of this study was cloning the Gba enzyme in pUCBM21 plasmid, and making frame mutation on it and sequencing it.Materials and methods: mRNA was extracted from mouse spleen and glucocerebrosidase cDNA was synthesized and amplified by PCR with specific primers. cDNA was cloned in pUCBM21 and analyzed by restriction enzymes. A fragment of its sequence was deleted using MscI restriction enzyme .Then frame mutation was prepared and sequenced.Results: glucocerebrosidase cDNA was synthesized and amplified by PCR. Then cloned in pUCBM21 and confirmed using restriction analysis. An in frame mutation was done and confirmed by sequencing.Conclusion: The successful sequenced Gba cDNA compared with other researchers results.

Aim and Background: Chemical mutagen mutagenic effects, diverse gene pool in plant resources and as a complementary tool in breeding programs are used. In this study, in order to make changes and changes in the morphological characteristics of different concentrations of active ingredients in the plant peppermint ethyl methane sulfonate were used EMS in vitro and field.Material and Methods: Research in vitro and farm with 4 treatments and 4 replications in a completely randomized design. From different treatments EMS includes 0/0% (control group), 0.05% and 0.01% and 0.005% and for different times (24 and 48) and the field was used in vitro. Evaluation was conducted in a test tube and simultaneously farm. After the transfer and development of subjects in conditions in vitro, treatment with the mutagen ethyl methane sulfonate were studied.Conclusion: The results showed that the use of MS medium mutagenic substances in medicinal plants play a role in phenotypic variation. Concentration of 0.01% EMS best result was significant at 1% and cultured explants were obtained, including that on the lateral buds and trailing characters such as stem length, number of leaves and buds of the side effect. The result analysis showed that the ethyl methane sulfonate oil treatments on materials essential components have not been affected.Results: Use mutagen EMS in peppermint plant tissue culture and in both instances treated plants on a farm stimulate the rapid growth of lateral buds of the plant, which increases the level of plant and thus increase the yield. Concentrations of greater than 0.05% EMS at 24 and 48 hours in loss of tissue samples and the field.

Dextransucrase of Leuconostoc mesenteroides PTCC 1059 is an enzyme of industrial and medical interest that catalyzes the synthesis of a soluble dextran from sucrose. The mutant strains of Leuconostoc mesenteroides PTCC 1059 hyperproducing for dextransucrase were isolated after UV irradiation and treatment with ethyl methane sulfonate. The enzyme activity of one mutant strain, the 1059M5E4, was about 2.5-fold higher than that of the wild type, while its cell growth was relatively lower. The 1059M5E4 dextransucrase produced the same type of dextran as well as the wild type but showed higher thermal stability. These properties may be interesting for using this strain in enzymatic production of dextran.