• Reproductive and Developmental Biology
• /
• 제32권4호
• /
• pp.255-261
• /
• 2008

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

• Asian-Australasian Journal of Animal Sciences
• /
• 제29권9호
• /
• pp.1239-1246
• /
• 2016

The breeding of yaks is highly seasonal, there are many crucial proteins involved in the reproduction control program, especially in follicular development. In order to isolate differential proteins between mature and immature follicular fluid (FF) of yak, the FF from yak follicles with different sizes were sampled respectively, and two-dimensional gel electrophoresis (2-DE) of the proteins was carried out. After silver staining, the Image Master 2D platinum software was used for protein analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was performed for differential protein identification. The expression level of transferrin and enolase superfamily member 1 (ENOSF1) was determined by Western blotting for verification analysis. The results showed that 2-DE obtained an electrophoresis map of proteins from mature and immature yak FF with high resolution and repeatability. A comparison of protein profiles identified 12 differently expressed proteins, out of which 10 of them were upregulated while 2 were downregulated. Western blotting showed that the expression of transferrin and ENOSF1 was enhanced with follicular development. Both the obtained protein profiles and the differently expressed proteins identified in this study provided experimental data related to follicular development during yak breeding seasons. This study also laid the foundation for understanding the microenvironment during oocyte development.

• Asian-Australasian Journal of Animal Sciences
• /
• 제25권11호
• /
• pp.1540-1545
• /
• 2012

Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism, but the profile of proteins associated with progesterone synthesis in cyclic and pregnant corpus luteum (CL) is not well-known in cattle. In Experiment 1, plasma progesterone level was monitored in cyclic cows (n = 5) and pregnant cows (n = 6; until d-90). A significant decline in the plasma progesterone level occurred at d-19 of cyclic cows. Progesterone level in abbatoir-derived luteal tissues was also determined at d 1 to 5, 6 to 13 and 14 to 20 of cyclic cows, and d-60 and -90 of pregnant cows (n = 5 each). Progesterone level in d-60 CL was not different from those in d 6 to 13 CL and d-90 CL, although the difference between d 6 to 13 and d-90 was significant. In Experiment 2, protein expression pattern in CL at d-90 (n = 4) was compared with that in CL of cyclic cows at d 6 to 13 (n = 5). Significant changes in the level of protein expression were detected in 32 protein spots by two-dimensional polyacrylamide gel electrophoresis (2-DE), and 23 of them were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Six proteins were found only in pregnant CL, while the other 17 proteins were found only in cyclic CL. Among the above 6 proteins, vimentin which is involved in the regulation of post-implantation development was included. Thus, the protein expression pattern in CL was disorientated from cyclic luteal phase to mid pregnancy, and alterations in specific CL protein expression may contribute to the maintenance of pregnancy in Korean native cows.

• 한국작물학회지
• /
• 제64권4호
• /
• pp.373-383
• /
• 2019

야생 호밀 염색체 첨가 계통의 단백질 발현 양상을 보통 밀과 비교함으로써 발현의 차이를 보이는 단백질의 기능을 동정함으로써 야생 호밀의 작물학적 유용 가치를 확인하고자 이 연구를 수행하였다. 전반적으로 야생 호밀 염색체 첨가계통은 보통 밀의 유전적 배경을 바탕으로 건조와 열에 대한 비생물학적 스트레스에 대한 저항성 관련 단백질과 바이러스성 병원균에 대한 저항성 관련 단백질 및 척박한 환경에 적응하는 생리대사에 관련된 단백질을 가지고 있는 것을 확인하였다. 하지만 아직 야생 호밀의 단백질 기능에 대한 정보와 작물학적 이용에 대한 연구가 미흡한 상태이다. 앞으로 국내 야생 호밀의 유용 유전자원으로써의 작물학적 이용과 기능에 대한 지속적인 연구가 필요하다.

• 환경생물
• /
• 제39권4호
• /
• pp.445-451
• /
• 2021

Agrobacterium tumefaciens strain CP4 유래 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) 유전자를 포함하는 유전자변형생물체(Living modified organism, LMO)가 개발되었다. 이 같은 LMO는 국내 승인되어 사료용, 식품용, 가공용으로 이용 중이다. 간이면역 검사키트 개발을 위해서는 고효율의 단클론 항체 개발이 필수적이다. 본 연구에서는 대장균 BL21 (DE3)에서 재조합 CP4 EPSPS 단백질을 정제하였으며 SDS-PAGE와 MALDI-TOF MS 분석으로 단백질 특성을 분석하였다. 단클론 항체 제작은 (주)앱클론의 SOP 매뉴얼에 따라 진행하였다. 본 연구 결과 5개의 단클론 항체 클론(2F2, 4B9, 6C11, 10A9, 10G9)를 확보하였다. 5종의 단클론 항체의 효율과 특이도 검정을 위해서 LM 면화 추출액을 이용한 western blotting 분석을 실시하였다. 모든 단클론 항체는 CP4 EPSPS를 함유하는 MON1445와 MON88913을 특이적으로 검출하였으며 비변형 면화 및 타종의 LM 면화에서는 검출되지 않았다. 이러한 결과들을 바탕으로 CP4 EPSPS 단클론 항체는 LMO에 함유된 CP4 EPSPS 단백질을 타겟으로 항체 기반 검출법 개발에 활용될 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권7호
• /
• pp.2951-2957
• /
• 2014

Background: Gastric carcinogenesis is a complicated process that involves environmental and genetic factors like interleukin-4 (IL-4) and IL-8. Single nucleotide polymorphisms in their genes are associated with changed levels of gene expression. Here, we investigated the association between IL4-590 C>T and IL8-251T>A and gastric cancer (GC) risk in Sichuan of Southwestern China. Materials and Methods: We surveyed the research subjects using a self-designed questionnaire with questions on demographic factors and putative risk factors. Approximately 2-5ml of whole blood was collected after field survey to analyze IL4-590 C>T and IL8-251T>A genotypes using MALDI-TOF MS. Results: Our study recruited 308 pairs of GC patients and controls, including 224 (72.7%) men and 84 (27.3%) women in each group. There were 99 cardia and 176 noncardia GC patients in the case group. The case and control groups had an average age of $57.7{\pm}10.6$ ($mean{\pm}SD$) and $57.6{\pm}11.1$ years. GC patients reported a significantly greater proportion of family history of cancer (29.9% vs 10.7%, p<0.01) and drinking (54.6% vs 43.2%, p<0.01) than did controls. Variant genotypes of IL-4-590 C>T and IL-8-251 T>A were not associated with overall GC risk (adjusted OR, 0.89; 95%CI, 0.61-1.28 for CT or CC vs TT; adjusted OR, 1.14; 95%CI, 0.86-1.79 for TA or AA vs TT). Stratification analysis of two SNPs for risk by subsites only found that variant IL-8-251 TA or AA genotype was associated with increased noncardia GC risk (adjusted OR, 2.58; 95%CI, 1.19-5.57). We did not observe interactions between the IL-8-251 T>A genotype and smoking (adjusted OR, 0.38; 95%CI, 0.08-1.79) or drinking (adjusted OR, 0.36; 95%CI, 0.08-1.65) for risk of noncardia GC. Conclusions: Our data indicate no association between the two SNPs of IL-4-590 and IL-8-251 with overall GC risk, while the IL-8-251 TA or AA genotype conferred risk of cardia GC. Our findings contribute to the evidence body for risk of SNPs associated with the development of gastric cancer in this region.