• 생명과학회지
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• 제28권4호
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• pp.389-397
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• 2018

당 단백질에 붙어 있는 당사슬 구조는 형질전환 돼지 유즙으로 분비되는 의약용 단백질의 생물학적 활성, 안정성 그리고 안전성에 영향을 줄 수 있다. 형질전환 동물을 이용한 치료용 당 단백질 생산은 유선 세포에서 이루어지는 당사슬 부가능력에 의해 제한되며, 균일한 당사슬 형태를 가지는 당 단백질 생산은 도전 과제로 남아있다. ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) 유전자는 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 단백질 당화기작에 중요한 효소이지만, 돼지 당 전이효소에 대한 정보는 매우 제한적이다. 따라서, 돼지 B3GNT1 (pB3GNT1) 유전자를 클로닝하고 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 기능적 특성을 조사하였다. 몇가지 다른 프라이머를 사용하여 전체 전사영역(ORF)을 함유하는 부분적인 pB3GNT1 mRNA 염기서열을 간 조직으로부터 분리하였다. 클로닝 된 pB3GNT1의 ORF는 1,248개의 뉴클레오티드를 가지며, 415개 아미노산 잔기로 구성되어 있었다. pB3GNT1 유전자의 장기별 발현특성은 성돈 및 자돈의 여러 기관에서 분석하였다. pB3GNT1 mRNA 발현 수준은 심장, 소장 보다는 근육에서 높았지만 폐에서는 낮았다. pB3GNT1의 기능적 특성 분석을 위해 돼지 신장 세포주(PK-15)에서 pB3GNT1 유전자의 안정적인 발현을 확립하였다. 그 결과, PK-15 세포에서 pB3GNT1 발현에 의한 당화 패턴은 총 시알산 증가에는 영향을 미치지 않지만, poly-N-아세틸글루코사민은 증가하는 것으로 나타났다. 본 연구는 생물반응기로 형질전환 돼지를 이용할 때 희망하는 당사슬을 부가하여 치료 가능성을 높이며 개선된 활성을 나타내는 당단백질 생산에 도움이 될 것이다.

• Molecules and Cells
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• 제22권3호
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• pp.343-352
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• 2006

Sialic acid is a sugar typically found at the N-glycan termini of glycoproteins in mammalian cells. Lec3 CHO cell mutants are deficient in epimerase activity, due to a defect in the gene that encodes a bifunctional UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE). Sialic acid modification on the cell surface is partially affected in these cells. We have mutagenized Lec3 CHO cells and isolated six mutants (termed C2m) deficient in the cell surface expression of polysialic acid (PSA). Mutant C2m9 was partially defective in expression of cell-surface PSA and wheat germ agglutinin (WGA) binding, while in the other five mutants, both cell-surface PSA and WGA binding were undetectable. PSA expression was restored by complementation with the gene encoding the CMP-sialic acid transporter (CST), indicating that CST mutations were responsible for the phenotypes of the C2m cells. We characterized the CST mutations in these cells by Northern blotting and RT-PCR. C2m9 and C2m45 carried missense mutations resulting in glycine to glutamate substitutions at amino acids 217 (G217E) and 256 (G256E), respectively. C2m13, C2m39 and C2m31 had nonsense mutations that resulted in decreased CST mRNA stability, and C2m34 carried a putative splice site mutation. PSA and CD15s expression in CST-deficient Lec2 cells were partially rescued by G217E CST, but not by G256E CST, although both proteins were expressed at similar levels, and localized to the Golgi. These results indicate that the novel missense mutations isolated in this study affect CST activity.

• Animal Bioscience
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• 제37권6호
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• pp.982-992
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• 2024

Objective: Jining Grey goat is a local Chinese goat breed that is well known for its high fertility and excellent meat quality but shows low meat production performance. Numerous studies have focused on revealing the genetic mechanism of its high fertility, but its highlighting meat quality and muscle growth mechanism still need to be studied. Methods: In this research, an integrative analysis of the genomics and transcriptomics of Jining Grey goats compared with Boer goats was performed to identify candidate genes and pathways related to the mechanisms of meat quality and muscle development. Results: Our results overlap among five genes (ABHD2, FN1, PGM2L1, PRKAG3, RAVER2) and detected a set of candidate genes associated with fatty acid metabolism (PRKAG3, HADHB, FASN, ACADM), amino acid metabolism (KMT2C, PLOD3, NSD2, SETDB1, STT3B, MAN1A2, BCKDHB, NAT8L, P4HA3) and muscle development (MSTN, PPARGC1A, ANKRD2). Several pathways have also been detected, such as the FoxO signaling pathway and Apelin signaling pathway that play roles in lipid metabolism, lysine degradation, N-glycan biosynthesis, valine, leucine and isoleucine degradation that involving with amino acid metabolism. Conclusion: The comparative genomic and transcriptomic analysis of Jining Grey goat and Boer goat revealed the mechanisms underlying the meat quality and meat productive performance of goats. These results provide valuable information for future breeding of goats.