Mi-Jeong Byun

Full-length enriched cDNA library construction from tissues related to energy metabolism in pigs

Genome sequencing of the pig is being accelerated because of its importance as an evolutionary and biomedical model animal as well as a major livestock animal. However, information on expressed porcine genes is insufficient to allow annotation and use of the genomic information. A series of expressed sequence tags of 5′ ends of five full-length enriched cDNA libraries (SUSFLECKs) were functionally characterized. SUSFLECKs were constructed from porcine abdominal fat, induced fat cells, loin muscle, liver, and pituitary gland, and were composed of non-normalized and normalized libraries. A total of 55,658 ESTs that were sequenced once from the 5′ ends of clones were produced and assembled into 17,684 unique sequences with 7,736 contigs and 9,948 singletons. In Gene Ontology analysis, two significant biological process leaf nodes were found: gluconeogenesis and translation elongation. In functional domain analysis based on the Pfam database, the beta transducin repeat domain of WD40 protein was the most frequently occurring domain. Twelve genes, including SLC25A6, EEF1G, EEF1A1, COX1, ACTA1, SLA, and ANXA2, were significantly more abundant in fat tissues than in loin muscle, liver, and pituitary gland in the SUSFLECKs. These characteristics of SUSFLECKs determined by EST analysis can provide important insight to discover the functional pathways in gene networks and to expand our understanding of energy metabolism in the pig.

Functional Analysis of Expressed Sequence Tags from Hanwoo (Korean Cattle) cDNA Libraries

임다정*변미정*조용민*윤두학*이승환*신윤희**임석기*농촌진흥청 축산과학원 생명환경부*, 인실리코젠**적 요본 연구는 한우의 지방, 간, 등심조직에서 유전자 염기서열을 확보하여 생산된 57,598개의 유전자 발현단편 데이터의 기능규명을 실시하였다. 유전자 발현단편 서열은 Assembly 과정을 통하여 unique한 서열인 4,759 contigs와 7,587 singletons을 확보하였으며, 얻어진 전사체를 이용하여 NCBI의 non-redundant 단백질 데이터베이스에 대하여 서열유사성 검색(BLAST)을 하여 유전자의 기능을 예측할 수 있었다. 또한 기능에 대한 모호성을 확실히 하기 위해 Gene Ontology 용어를 사용하여 한우의 세 조직에서 확보된 서열들의 생물학적 특성을 기술하였다. Gene Ontology는 모든 기능이 계층적으로 표현되어 있기 때문에, 각 계층에 대하여 유의적인 기능 여부를 확인하기 위하여 통계분석인 Pearson’s chi-square test를 실시하여 통계적으로 유의한 기능들을 산출할 수 있었다. 그 결과, Molecular function, Biological process, Cellular component 각각의 GO category에서 13, 16, 8개의 유의적인 GO terms이 검출되었다. 또한, 한우의 세 조직에 대하여 조직특이적 유전자의 존재여부를 판단하기 위하여 Audic’s test를 실시하여 세 조직에서 각각 조직특이적으로 발현되는 유전자들을 검출할 수 있었다. 이러한 생물정보학적 방법들을 사용하여 한우의 세 조직에서 발현된 대량의 서열들에 대한 기능을 예측할 수 있었으며, 통계 검증을 통하여 유의적으로 검출된 유전자들은 추후에 실험적 검증을 실시하여 충분한 정보를 확보할 수 있을 것으로 사료된다.(Key words :Hanwoo, EST, Gene Ontology, Audic’s test)

Effects of Kinds of Cryoprotectants on the Characteristics of Frozen Fowl Semen

Department of Animal Science, Division Applied Life Institute Agriculture and Graduate School Gyeongsang National University, Jinju 660-701, KoreaABSTRACT The purpose of this study was to evaluate the sperm viability, normal acrosome and mitochondrial activity in the frozen-thawed fowl semen by different cryoprotectants. The experiment was carried out on 10 sexually adult roosters of Ogye. The semen was collected twice a week and pooled semen was diluted 1:1 EK extender containing no cryoprotectant at 5. After equilibration for 30 minutes, diluted chicken semen was diluted 1:1 extender containing either 7% dimethyl- acetamide (DMA), 7% dimethylformamide (DMF) or 7.5% methylacetamide (MA) at final concentration and was put in 0.5 mL plastic straws and frozen for 30 minutes by exposure to liquid nitrogen vapor 4 cm above the surface of liquid nitrogen, followed by plunging into liquid nitrogen. Frozen semen was thawed in water bath at 5for 2 minutes. For cytometric analysis, the frozen-thawed semen was diluted with EK extender to a final concentration of 90 million spermatozoa per mL. Sperm membrane integrity was evaluated as SYBR-14 and propidium iodide (PI). Acrosome integrity was assessed with fluorescein isothiocyanate-labeled PSA and PI. The percentage of mitochondrial function was estimated by using Rhodamine123 (R123) and PI. In conclusion, freezing rooster semen by using 7% DMF as cryoprotectant was significantly highest in rates of survival and mitochondrial function while its rate of damage of acrosome was significantly lowest. As a result, DMF is the cryoprotectant that has the lowest influences on sperm membranes and acrosome integrity. Therefore it could be used for freezing method of animal genetic conservation method for poultry diversity.(Key words : semen, FACS, Dimethylacetamide, Dimethylformamide, N-Methylacetamide)

Comparison of Growth Performance between Korean Native Layer Chickens and Imported Layer Chickens at Early Rearing Stage

Department of Animal Science, Chonnam National UniversityABSTRACT The objective of this study was to compare the growth performance between Korean native layer chickens and imported layer chickens at early rearing stage. Total number of chicks analyzed in this study was 276 and feeding period was conducted from July 24, 2012 for 10 weeks. Five strains including 2 Korean native strains: A=Korean Native Black (Chung-cheongbuk-do) and B=Korean Native Yellowish Brown (Gyeongsangbuk-do) and 3 imported layer strains: C=White Leghorn (Gyeongsangnam-do), D=White Leghorn (Seoul), and E=Ameraucanas (Gyeongsangbuk-do) were used to analyze the following traits such as fertility, hatchability, body weight at a different growing stage, average body weight gain, and feed conversion ratio. The fertilities and hatchabilities of strains were 93.88% and 95.65% in strain A, 81.75% and 86.24% in strain B, 82.25% and 88.15% in strain C, 79.25% and 90.85% in strain D, and 71.50% and 88.11% in strain E, respectively. A viability was excellent in strains A and E to be more than 98% and was low in strain D to be 86.67% at a whole week. The strain A had greater body weight during growing stages (p<0.05) than the other strains. The shank length of strain D of 56.69±3.27 mm was the highest value at 10 weeks of age among strains (p<0.05). The phenotypic correlation coefficients of strains A and D between an average body weight gain and a shank length were 0.63 and 0.73 during 02 wk, 0.70 and 0.55 during 24 wk, 0.55 and 0.54 during 46 wk, 0.50 and 0.24 during 68 wk, and 0.46 and 0.29 during 810 wk, respectively. The Korean native hens may have potential abilities to be used as an excellent seed stock for poultry industry.(Key words : Korean native chickens, imported layer chickens, early rearing stage, body weight, shank length)

Effect of N-Methylacetamide Concentration on the Fertility and Hatchability of Cryopreserved Ogye Rooster Semen

Poutry Science Division, National Institute of Animal Science, RDA, Seonghwan 330-801, KoreaABSTRACT To preserve chicken genetic materials like cryopreserved spermatozoa, various kinds of freezing agents like glycerol, dimethylsuloxide, dimethylformamide or dimethylacetamide have been used for rooster semen preparation. Recently, the usage of N-methylacetamide (MA) for Ogye rooster semen preservation resulted in hatched chicken successfully. In this study, we investigated the effects of 7, 9 and 11% of MA on the viability, fertility and hatchability of frozen-thawed rooster semen using artificial insemination. The results of viability, fertility and hatchability in frozen semen with 7%, 9% or 11% MA were 35.16±6.12%, 67.83±15.3% and 66.2±16.3% of motile sperm rate, 21.5%, 34.7% and 25% of fertility rate, and 100%, 89.5% and 87.5% of hatchability rate. The results of control group with frozen semen were 96.0% of fertility rate and 92.2% of hatchability rate. With these results, the concentration range of MA as a freezing agent of rooster semen could be 79% of media. The higher concentration of 9 % MA could decrease the fertility rate of thawed semen not the rate of hatchability rate. So the use of MA without affecting fertility rate would be a key point of freezing method of rooster semen for poultry genetic resource preservation.(Key words : Ogye, cryopreservation, N-methylacetamide, semen)

Analysis of Genetic Diversity and Relationships of Korean Native Goat Populations by Microsatellite Markers

본 연구는 30개의 MS 마커를 이용하여 한국재래염소 3개 집단(당진, 장수, 통영)과 1개 농가집단을 대상으로 집단 내 및 집단간의 유전적 다양성, 계통유전학적 유연관계 분석 및 한국재래염소 3개 집단간의 유전적 균일성을 검증하여 우리 고유유전자원으로서의 가치를 구명하고자 실시하였다. 대립유전자형 분석 결과, 총...