- Email: [email protected]
Effects of day 0 protocol for superovulation on embryo yields and ovarian responses in Kilis and Angora goats
Abstracts / Current Opinion in Biotechnology 22S (2011) S15–S152
of estrous and embryos transfer as efficient modalities to use the valuable genetic material. doi:10.1016/j.copbio.2011.05.147
S55
are now to discovered further with bioinformatic tools to be available either as sequence-tagged sites (STS) for these organisms having little molecular information or for further real-time RAPD SNP or polymorphism detections. doi:10.1016/j.copbio.2011.05.149
A15 PLS-DA approach for simultaneous evaluation of genes involved in luteolysis in equine endometrium during estrous cycle and early pregnancy Guzeloglu 1 ,
Kayis 1 ,
Aydin Seyit A Ahmet Semacan 1 , Selim Aslan 3
Mehmet O
Atli 2 ,
Ercan
Kurar 1 ,
1
Selcuk University, Konya, Turkey Dicle University, Diyarbakır, Turkey 3 Ankara University, Ankara, Turkey 2
E-mail address: [email protected] (A. Guzeloglu) Expression of genes involved in luteolysis is tightly regulated by cyclic changes in hormonal levels and embryonic presence in the uterus. Inhibition of luteolysis is vital for the maintenance of pregnancy real-time PCR studies evaluate each gene separately. Aims of this study were to (1) analyze expression of genes simultaneously and (2) obtain degree of contribution of genes to separation of treatment groups. Therefore, partial least square regression discriminant analysis (PLS-DA), characterized by high computational and statistical efficiency, was employed. Biopsies obtained from mares in following treatment groups; late diestrus (LD, n = 4), early luteolysis (EL, n = 4) and estrus phase (AL, n = 4) of the cycle and days 14 (P14; n = 4), 15 (P15, n = 4), and 18 (P18, n = 4) in pregnant groups. Relative mRNA expression levels of genes were quantified using real-time RT-PCR. Treatment groups were separated efficiently via the PLS-DA. First three components of the predictor score variables explained 73.2% of information of 12 genes (t1 = 40.79%, t2 = 20.91%, and t3 = 11.50%). PLS-DA method can be used to determine the degree of contribution of each gene in a physiological event examined. Acknowledgements: Funded by TUBITAK-TOVAG 107O035. doi:10.1016/j.copbio.2011.05.148
A17 Effects of day 0 protocol for superovulation on embryo yields and ovarian responses in Kilis and Angora goats Ali Reha Agaoglu 1 , Mustafa Kaymaz 2 , Umut Tasdemir 3 1
Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Mehmet Akif Ersoy University, Burdur, Turkey 2 Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Ankara University, Ankara, Turkey 3 Ministry of Agriculture and Rural Affairs, Lalahan Livestock Central Research Institute, Ankara, Turkey E-mail address: [email protected] (A.R. Agaoglu) The aim of the study was to assess the effects of day 0 protocol for superovulation treatment on embryo yields and ovarian responses in Angora and Kilis goats in the out of breeding season. 16 Angora and 11 Kilis goats were used in this study. After controlled internal drug release withdrawal estrus signs were observed in the superovulation process. Then natural mating was performed. Embryos were collected and number of corpora lutea were recorded with laparotomy. Transferable embryo yields were significantly higher in Angora goats (31/42, 74%) compared to Kilis goats (16/46, 35%) in the non-breeding season (P < 0.01): the day 0 protocol for superovulation can be used in goats in the out of breeding season. Whereas transferable embryo yields are affected by the different goat breed. A lower number of transferable embryos that were obtained from Kilis goats in the present study might be due to fertilization failure and early regression of corpus luteum. This study was supported by the Scientific and Technological Research Council of Turkey (TUBITAK) (project no. 106G105, TURKHAYGEN-1). doi:10.1016/j.copbio.2011.05.150
A16 A downhill strategy for RAPD and real-time RAPD derived SCAR markers Esma Banu Ozsoy, Zeynep Katmer, Tugba Senel, Irem Uzonur Biology Department, Faculty of Sciences and Engineering, Fatih University, Istanbul, Turkey E-mail address: [email protected] (E.B. Ozsoy) RAPD-PCR has been widely used for locus-specific SCAR marker production. However, it is not that convenient to produce such markers with traditional and widely accepted methods starting with the isolation of RAPD band from the gel, followed with a reamplification step and then cloning of the product and sequencing it to produce the new longer and specific primers for the amplification of required DNA sequences in downstream applications. In our work we have developed a downhill strategy for SCAR marker production. RAPD profiles for the two world-wide important mariculture species Sparus auratus and Dicentrarchus labrax using different combinations of two RAPD primers have been produced and the band(s) that were confirmed to be amplified with both RAPD primers were isolated and directly sequenced. Primers were designed to specifically amplify anonymous sequences which
Biocatalysis & Biotransformation Section B1 The effects of pH and temperature on the production of antioxidant enzymes by Rhodotorula glutinis Ayse Ezgi Unlu, Serpil Takac Department of Chemical Engineering, Ankara University, Ankara, Turkey E-mail address: [email protected] (A.E. Unlu) Organisms need oxygen to maintain vital requirements. However, various detrimental effects and pollutants cause oxygen to produce toxic and lethal intermediates. Organisms protect themselves with non-enzymatic and enzymatic antioxidants. The two important components of antioxidants are superoxide dismutase (SOD) and catalase (CAT) enzymes. The aim of this work is to investigate the effects of pH and temperature on the production SOD and CAT, by a high antioxidant producing yeast, R. glutinis. The yeast was grown in UYM culture medium (30◦ C, 150 rpm, 96 h). The effects of incubation temperature (10–35◦ C) and initial pH
of estrous and embryos transfer as efficient modalities to use the valuable genetic material. doi:10.1016/j.copbio.2011.05.147
S55
are now to discovered further with bioinformatic tools to be available either as sequence-tagged sites (STS) for these organisms having little molecular information or for further real-time RAPD SNP or polymorphism detections. doi:10.1016/j.copbio.2011.05.149
A15 PLS-DA approach for simultaneous evaluation of genes involved in luteolysis in equine endometrium during estrous cycle and early pregnancy Guzeloglu 1 ,
Kayis 1 ,
Aydin Seyit A Ahmet Semacan 1 , Selim Aslan 3
Mehmet O
Atli 2 ,
Ercan
Kurar 1 ,
1
Selcuk University, Konya, Turkey Dicle University, Diyarbakır, Turkey 3 Ankara University, Ankara, Turkey 2
E-mail address: [email protected] (A. Guzeloglu) Expression of genes involved in luteolysis is tightly regulated by cyclic changes in hormonal levels and embryonic presence in the uterus. Inhibition of luteolysis is vital for the maintenance of pregnancy real-time PCR studies evaluate each gene separately. Aims of this study were to (1) analyze expression of genes simultaneously and (2) obtain degree of contribution of genes to separation of treatment groups. Therefore, partial least square regression discriminant analysis (PLS-DA), characterized by high computational and statistical efficiency, was employed. Biopsies obtained from mares in following treatment groups; late diestrus (LD, n = 4), early luteolysis (EL, n = 4) and estrus phase (AL, n = 4) of the cycle and days 14 (P14; n = 4), 15 (P15, n = 4), and 18 (P18, n = 4) in pregnant groups. Relative mRNA expression levels of genes were quantified using real-time RT-PCR. Treatment groups were separated efficiently via the PLS-DA. First three components of the predictor score variables explained 73.2% of information of 12 genes (t1 = 40.79%, t2 = 20.91%, and t3 = 11.50%). PLS-DA method can be used to determine the degree of contribution of each gene in a physiological event examined. Acknowledgements: Funded by TUBITAK-TOVAG 107O035. doi:10.1016/j.copbio.2011.05.148
A17 Effects of day 0 protocol for superovulation on embryo yields and ovarian responses in Kilis and Angora goats Ali Reha Agaoglu 1 , Mustafa Kaymaz 2 , Umut Tasdemir 3 1
Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Mehmet Akif Ersoy University, Burdur, Turkey 2 Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Ankara University, Ankara, Turkey 3 Ministry of Agriculture and Rural Affairs, Lalahan Livestock Central Research Institute, Ankara, Turkey E-mail address: [email protected] (A.R. Agaoglu) The aim of the study was to assess the effects of day 0 protocol for superovulation treatment on embryo yields and ovarian responses in Angora and Kilis goats in the out of breeding season. 16 Angora and 11 Kilis goats were used in this study. After controlled internal drug release withdrawal estrus signs were observed in the superovulation process. Then natural mating was performed. Embryos were collected and number of corpora lutea were recorded with laparotomy. Transferable embryo yields were significantly higher in Angora goats (31/42, 74%) compared to Kilis goats (16/46, 35%) in the non-breeding season (P < 0.01): the day 0 protocol for superovulation can be used in goats in the out of breeding season. Whereas transferable embryo yields are affected by the different goat breed. A lower number of transferable embryos that were obtained from Kilis goats in the present study might be due to fertilization failure and early regression of corpus luteum. This study was supported by the Scientific and Technological Research Council of Turkey (TUBITAK) (project no. 106G105, TURKHAYGEN-1). doi:10.1016/j.copbio.2011.05.150
A16 A downhill strategy for RAPD and real-time RAPD derived SCAR markers Esma Banu Ozsoy, Zeynep Katmer, Tugba Senel, Irem Uzonur Biology Department, Faculty of Sciences and Engineering, Fatih University, Istanbul, Turkey E-mail address: [email protected] (E.B. Ozsoy) RAPD-PCR has been widely used for locus-specific SCAR marker production. However, it is not that convenient to produce such markers with traditional and widely accepted methods starting with the isolation of RAPD band from the gel, followed with a reamplification step and then cloning of the product and sequencing it to produce the new longer and specific primers for the amplification of required DNA sequences in downstream applications. In our work we have developed a downhill strategy for SCAR marker production. RAPD profiles for the two world-wide important mariculture species Sparus auratus and Dicentrarchus labrax using different combinations of two RAPD primers have been produced and the band(s) that were confirmed to be amplified with both RAPD primers were isolated and directly sequenced. Primers were designed to specifically amplify anonymous sequences which
Biocatalysis & Biotransformation Section B1 The effects of pH and temperature on the production of antioxidant enzymes by Rhodotorula glutinis Ayse Ezgi Unlu, Serpil Takac Department of Chemical Engineering, Ankara University, Ankara, Turkey E-mail address: [email protected] (A.E. Unlu) Organisms need oxygen to maintain vital requirements. However, various detrimental effects and pollutants cause oxygen to produce toxic and lethal intermediates. Organisms protect themselves with non-enzymatic and enzymatic antioxidants. The two important components of antioxidants are superoxide dismutase (SOD) and catalase (CAT) enzymes. The aim of this work is to investigate the effects of pH and temperature on the production SOD and CAT, by a high antioxidant producing yeast, R. glutinis. The yeast was grown in UYM culture medium (30◦ C, 150 rpm, 96 h). The effects of incubation temperature (10–35◦ C) and initial pH