• Asian-Australasian Journal of Animal Sciences
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• 제33권2호
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• pp.219-229
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• 2020

Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORT^TM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.

• Asian-Australasian Journal of Animal Sciences
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• 제33권10호
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• pp.1544-1557
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• 2020

Objective: Genomic selection (GS) is becoming popular in animals' genetic development. We, therefore, investigated the single-step genomic best linear unbiased prediction (ssGBLUP) as tool for GS, and compared its efficacy with the traditional pedigree BLUP (pedBLUP) method. Methods: A total of 9,952 males born between 1997 and 2018 under Hanwoo proven-bull selection program was studied. We analyzed body weight at 12 months and carcass weight (kg), backfat thickness, eye muscle area, and marbling score traits. About 7,387 bulls were genotyped using Illumina 50K BeadChip Arrays. Multiple-trait animal model analyses were performed using BLUPF90 software programs. Breeding value accuracy was calculated using two methods: i) Pearson's correlation of genomic estimated breeding value (GEBV) with EBV of all animals (r_M1) and ii) correlation using inverse of coefficient matrix from the mixed-model equations (r_M2). Then, we compared these accuracies by overall population, info-type (PHEN, phenotyped-only; GEN, genotyped-only; and PH+GEN, phenotyped and genotyped), and bull-types (YBULL, young male calves; CBULL, young candidate bulls; and PBULL, proven bulls). Results: The r_M1 estimates in the study were between 0.90 and 0.96 among five traits. The r_M1 estimates varied slightly by population and info-type, but noticeably by bull-type for traits. Generally average r_M2 estimates were much smaller than r_M1 (pedBLUP, 0.40 to0.44; ssGBLUP, 0.41 to 0.45) at population level. However, r_M2 from both BLUP models varied noticeably across info-types and bull-types. The ssGBLUP estimates of r_M2 in PHEN, GEN, and PH+ GEN ranged between 0.51 and 0.63, 0.66 and 0.70, and 0.68 and 0.73, respectively. In YBULL, CBULL, and PBULL, the r_M2 estimates ranged between 0.54 and 0.57, 0.55 and 0.62, and 0.70 and 0.74, respectively. The pedBLUP based r_M2 estimates were also relatively lower than ssGBLUP estimates. At the population level, we found an increase in accuracy by 2.0% to 4.5% among traits. Traits in PHEN were least influenced by ssGBLUP (0% to 2.0%), whereas the highest positive changes were in GEN (8.1% to 10.7%). PH+GEN also showed 6.5% to 8.5% increase in accuracy by ssGBLUP. However, the highest improvements were found in bull-types (YBULL, 21% to 35.7%; CBULL, 3.3% to 9.3%; PBULL, 2.8% to 6.1%). Conclusion: A noticeable improvement by ssGBLUP was observed in this study. Findings of differential responses to ssGBLUP by various bulls could assist in better selection decision making as well. We, therefore, suggest that ssGBLUP could be used for GS in Hanwoo proven-bull evaluation program.

• 미생물학회지
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• 제44권3호
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• pp.178-185
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• 2008

조류 독감바이러스(avian influenza virus, AIV)는 사람에게서 발생하는 인플루엔자 대유행에 중요한 역할을 한다. 특히 최근 AIV H9N2형에 의한 가금류 감염이 빈번히 나타나고 있어 인체 감염이 상당히 우려되는 실정이다. 본 연구에서는 최종적으로 AIV의 HA와 NA 단백질에 특이적으로 반응하는 항혈청을 생산하고자 하였다. 먼저 감염된 닭에서 분리된 AIV H9N2 한국분리주 A/Ck/Kr/MS96/96의 게놈RNA로부터 RT-PCR 방법으로 HA와 NA 단백질 N-말단부위에 해당하는 염기서열을 증폭하였다. 이렇게 증폭된 DNA단편은 E. coli 발현벡터 pGEX4T-1에 삽입한 후, BL21 세포에서 각각의 GST fusion protein (GST-HAln와 GST-NAn) 형태로 발현하였다. GST-HAln와 GST-NAn은 모두 glutathione sepharose column을 사용하여 분리 및 정제하였으며, 정제된 단배질을 항원으로 사용하여 토끼 항혈청을 생산하였다. 생산된 항혈청의 항원특이성은 AIV H9N2 한국분리주 A/Ck/Kr/MS96/96로 감염된 MDCK 세포의 cell extract를 사용하여 immunoblotting을 수행함으로써 확인하였다. 본 실험결과AIV H9N2의 HA와 NA단백질 N-말단부위에 해당하는 재조합GST fusion protein과, 이들 각각의 단백질에 특이적으로 반응하는 항혈청은 앞으로AIV 감염의 진단 뿐만 아니라, AIV에 대한 기초연구에 중요한 재료로 사용될 것으로 기대한다.

• 한국응용과학기술학회지
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• 제31권1호
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• pp.31-43
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• 2014

This experiment evaluated the interaction effect of extreme heat diet(EHD), inverse lighting, and cool water on the growth performance of broiler chickens under extreme heat stress. There were 4 experimental groups (T1: EHD 1, 10:00-19:00 dark, 19:00-10:00 light, cold water $9^{\circ}C$; T2: EHD 2, 10:00-19:00 dark, 19:00-10:00 light, cold water $9^{\circ}C$; T3: EHD 1, 09:00-18:00 dark, 18:00-09:00 light, cold water $14^{\circ}C$; T4: EHD 2, 09:00-18:00 dark, 18:00-09:00 light, cold water $14^{\circ}C$), each group composed of 25 broilers and the experiment was repeated 3 times. EHD 1 contained soybean oil, molasses, methionine and lysine. EHD 2 contained all nutrients of EHD 1 and vitamin C additionally. As a result, T1 and T2 displayed higher body weight increase and diet intake compared to T3 and T4 (p<0.05). The weights of their liver and gizzard were similar but the weights of the thymus and bursa F were higher for T1 and T2 compared to that of T3 and T4 (p<0.05). It was observed that T1 and T2 displayed higher concentrations of blood triglyceride, total cholesterol, HDL-C and blood sugar compared to that of T3 and T4 but LDL-C level was higher for T3 and T4 compared to that of T1 and T2 (p<0.05). T1 and T2 displayed higher levels of immunity substances such as IgG, IgA and IgM compared to T3 and T4 but the blood level of corticosterone displayed to be lower for T1 and T2 compared to T3 and T4 (p<0.05). The T1 and T2 contained a higher amount of fecal lactobacillus compared to that of T3 and T4 but the T3 and T4 contained a higher amount of fecal E. coli, total aerobic bacteria, coliform bacteria compared to that of T1 and T2 (p<0.05). T1 and T2 displayed higher concentrations of cecal acetic acid, propionic acid and total short chain fatty acids compared to T3 and T4 but T3 and T4 displayed higher concentrations of butyric acid, isobutyric acid, valeric acid and isovaleric acid compared to T1 and T2 (p<0.05). These results have been observed that broiler chickens exposed to extreme heat stress with feeding EHD, inverse lighting and cold water would improve blood lipid, and elevate the production of immunity substance, beneficial microorganisms, and short chain fatty acids. This provision would also reduce the blood sugar consumption rate as energy sources and these effects will improve the growth performance of the broilers exposed to extreme heat.