Expression of Biologically Active Human Interferon Gamma in the Milk of Transgenic Mice Under the Control of the Murine Whey Acidic Protein Gene Promoter

Bağış, Haydar; Aktoprakligil, Diğdem; Güneş, Çagatay; Arat, Sezen; Akkoç, Tolga; Çetinkaya, Gaye; Kankavi, Orhan; Taşkın, Ali Cihan; Arslan, Korhan; Dündar, Munis; Tsoncheva, Vania L.; Ivanov, Iván

doi:10.1007/s10528-010-9403-7

Cited by 12 publications

(5 citation statements)

References 18 publications

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“…Next, mammary glands were gently massaged and milked with tweezers. According to the method described previously 43 44 , milk collection was slightly modified and started on day 8 after delivery. Briefly, samples were collected 3 times each day, 100 μl each time, and continually for 5 days.…”

Section: Methodsmentioning

confidence: 99%

Intron V, not intron I of human thrombopoietin, improves expression in the milk of transgenic mice regulated by goat beta-casein promoter

Hao

Zhou

et al. 2015

Sci Rep

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Introns near 5′ end of genes generally enhance gene expression because of an enhancer /a promoter within their sequence or as intron-mediated enhancement. Surprisingly, our previous experiments found that the vector containing the last intron (intron V) of human thromobopoietin (hTPO) expressed higher hTPO in cos-1 cell than the vector containing intron I regulated by cytomegalovirus promoter. Moreover, regulated by 1.0 kb rat whey acidic protein promoter, hTPO expression was higher in transgenic mice generated by intron V-TPOcDNA than in transgenic mice generated by TPOcDNA and TPOgDNA. However, it is unknown whether the enhancement of hTPO expression by intron I is decreased by uAUG7 at 5′-UTR of hTPO in vivo. Currently, we constructed vectors regulated by stronger 6.5kb β-casein promoter, including pTPOGA (containing TPOcDNA), pTPOGB (containing TUR-TPOcDNA, TUR including exon1, intron I and non-coding exon2 of hTPO gene), pTPOGC (containing ΔTUR-TPOcDNA, nucleotides of TUR from uAUG7 to physiological AUG were deleted), pTPOGD (containing intron V-TPOcDNA) and pTPOGE (containing TPOgDNA), to evaluate the effect of intron I on hTPO expression and to further verify whether intron V enhances hTPO expression in the milk of transgenic mice. The results demonstrated that intron V, not intron I improved hTPO expression.

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Section: Methodsmentioning

confidence: 99%

Intron V, not intron I of human thrombopoietin, improves expression in the milk of transgenic mice regulated by goat beta-casein promoter

Hao

Zhou

et al. 2015

Sci Rep

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“…Among mammalian expression systems; CHO was the focal point of studies from the 1990s onward, achieving laboratory-scale secretion of 15 mg L À1 of hIFNg [28] and studies included overexpression, optimisation of cultivation, scale-up production and purification [3,6,29,30]. An order of magnitude greater expression yields of hIFNg (570 mg mL À1 ) was achieved in mammary glands of transgenic mice in vivo, which is comparable to productivity in E. coli [7,31]. Studies in mammalian expression systems are ongoing to improve productivities further and to lower the cost of production, which is essential to make mammalian expression systems at the industrial scale competitive with the currently used E. coli expression system.…”

Section: C(t)mentioning

confidence: 99%

“…To overcome these limitations, expression of recombinant hIFNg was attempted in various hosts like Saccharomyces cerevisiae 20B-12 [4], insect cells lines Spodoptera frugiperda, Spodoptera exigua, and Spodoptera litura. [5], Chinese hamster ovary (CHO) [3,6], wild-type mice strain C57BL/6 [7], rat cell line 3Y1-B [8], monkey and human cells [9]; however; high costs of cultivation and purification, contamination, low yields, low biological activity and short half-life of the product also adversely impacted on the use of these expression systems [10].…”

Section: Introductionmentioning

confidence: 99%

Is Pichia pastoris a realistic platform for industrial production of recombinant human interferon gamma?

et al. 2017

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Human interferon gamma (hIFNγ) is an important cytokine in the innate and adaptive immune system, produced commercially in Escherichia coli. Efficient expression of hIFNγ has been reported once for Pichia pastoris (Wang et al., 2014) - a proven heterologous expression system. This study investigated hIFNγ expression in P. pastoris replicating the previous study and expanding by using four different strains (X33: wild type; GS115: HISMut; KM71H: Arg, Mut and CBS7435: Mut) and three different vectors (pPICZαA, pPIC9 and pPpT4αS). In addition, the native sequence (NS) and two codon-optimised sequences (COS1 and COS2) for P. pastoris were used. Methanol induction yielded no expression/secretion of hIFNγ in X33, highest levels were recorded for CBS7435: Mut (∼16 μg. L). mRNA copy number calculations acquired from RT-qPCR for GS115-pPIC9-COS1 proved low abundance of mRNA. A 10-fold increase in expression of hIFNγ was achieved by lowering the minimal free energy of the mRNA and 100-fold by Mut phenotypes, substantially lower than reported by Wang et al. (2014). We conclude that commercial production of low cost, eukaryotic recombinant hIFNγ is not an economically viable in P. pastoris. Further research is required to unravel the cause of low expression in P. pastoris to achieve economic viability.

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“…There are variations among different mouse strains in terms of in vitro development rates (13)(14), embryonic stem cell obtaining rates (15), and also cloning success ratios. B6CBAF1 (C57BL/6j×CBA/J), C57BL/6j, and B6D2F1 (C57BL/6j×DBA/J) mouse strains are common used in reproductive biotechnology (ICSI, SCNT and transgenic model) (16)(17)(18). No research was found in literature about comparative results of in vitro developments among oocytes belonging to these strains mouse.…”

Section: Introductionmentioning

confidence: 99%

Comparison of parthenogenetic oocyte activation in different mouse strains on in vitro development rate and quality

Taşkın¹,

Kocabay²,

Gül³

et al. 2021

Veteriner Hekimler Derneği Dergisi

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Çalışmamızın amacı, partenogenetik aktivasyonda farklı fare ırklarında in vitro embriyo gelişimi ve kalitesi üzerindeki etkilerinin araştırılmasıdır. Bu çalışmada, B6CBAF1, C57BL/6j, and B6D2F1 farelerin superovulasyon ile elde edilen oositleri kullanılmıştır. Superovule edilen fareler, insan koryonik gonadotropin (hCG) uygulamasından 14 saat sonra oositler elde edildi ve 18 saat sonra partenogenetik aktivasyona başlandı. Oositler, 10 mM SrCl2 + 5 μg/mL-1 sitokalazin B (CB) + 5 nM trikostatin A (TSA) Ca 2+ içermeyen Chatot Ziomek Brinster (CZB) medyumu içerisinde 6 saat bekletildi. Aktivasyon sonrası, embriyo kültür medyumu + TSA’da inkübatörde 37°C ve %5 CO2 ortamında 2 saat bekletildi. Son olarak, tüm embriyolar 120 saat süre ile kültüre edildi. Bu çalışmadan elde edilen sonuçlar göre, B6D2F1 ırkının partenogenetik aktivasyon başarısı, C57BL/6j ve B6CBAF1 ırklarına göre daha yüksek bulundu.

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Expression of Biologically Active Human Interferon Gamma in the Milk of Transgenic Mice Under the Control of the Murine Whey Acidic Protein Gene Promoter

Cited by 12 publications

References 18 publications

Intron V, not intron I of human thrombopoietin, improves expression in the milk of transgenic mice regulated by goat beta-casein promoter

Intron V, not intron I of human thrombopoietin, improves expression in the milk of transgenic mice regulated by goat beta-casein promoter

Is Pichia pastoris a realistic platform for industrial production of recombinant human interferon gamma?

Comparison of parthenogenetic oocyte activation in different mouse strains on in vitro development rate and quality

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