Construction of a recombinant human insulin expression vector for mammary gland-specific expression in buffalo (Bubalus bubalis) mammary epithelial cell line

Kaushik, Ravinder; Singh, Kiran; Kumari, Archana; Rameshbabu, K.; Singh, Manoj Kumar; Manik, R. S.; Palta, P.; Singla, S. K.; Chauhan, M. S.

doi:10.1007/s11033-014-3464-3

Cited by 7 publications

(5 citation statements)

References 51 publications

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“…Most of the plasmid vectors are from 3 kbp to 5 kbp in length. Plasmids allow inserting a sequence of DNA up to 20 kbp in length, and have been used to obtain transgenic animals (Kaushik et al 2014). The commercial vector pBC1 with a DNA of 21.6 kbp long has been also used to obtain transgenic cattle (Yang et al 2011).…”

Section: Vectors Used In the Creation Of Transgenic Animalsmentioning

confidence: 99%

“…In 1998, Ilan et al (1998 suggested that MEC cultures could be an alternative to transgenic mice models to assess the poten-tial gene constructs for expression in the mammary gland of transgenic farm animals. Some other studies have used MEC cultures to assess the expression of the established vectors (Monzani et al 2011;Monzani et al 2013;Kaushik et al 2014;Monzani et al 2015). Although, transgenic mice can be the best model for transgenic farm animals, MEC culture is a strategy that is easier to implement than the creation of transgenic mice.…”

Section: The Evaluation Of the Constructed Vectorsmentioning

confidence: 99%

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Genetically modified animals for use in biopharmacology: from research to production

Deykin¹,

Shcheblykina²,

Povetka³

et al. 2021

RRP

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Introduction: In this review, the analysis of technologies for obtaining biologically active proteins from various sources is carried out, and the comparative analysis of technologies for creating producers of biologically active proteins is presented. Special attention is paid to genetically modified animals as bioreactors for the pharmaceutical industry of a new type. The necessity of improving the technology of development transgenic rabbit producers and creating a platform solution for the production of biological products is substantiated. The advantages of using TrB for the production of recombinant proteins: The main advantages of using TrB are the low cost of obtaining valuable complex therapeutic human proteins in readily accessible fluids, their greater safety relative to proteins isolated directly from human blood, and the greater safety of the activity of the native protein. The advantages of the mammary gland as a system for the expression of recombinant proteins: The mammary gland is the organ of choice for the expression of valuable recombinant proteins because milk is easy to collect in large volumes. Methods for obtaining transgenic animals: The modern understanding of the regulation of gene expression and the discovery of new tools for gene editing can increase the efficiency of creating bioreactors for animals and help to obtain high concentrations of the target protein. The advantages of using rabbits as bioreactors producing recombinant proteins in milk: The rabbit is a relatively small animal with a short duration of gestation, puberty and optimal size, capable of producing up to 5 liters of milk per year per female, receiving up to 300 grams of the target protein.

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Section: Vectors Used In the Creation Of Transgenic Animalsmentioning

confidence: 99%

Section: The Evaluation Of the Constructed Vectorsmentioning

confidence: 99%

Genetically modified animals for use in biopharmacology: from research to production

Deykin¹,

Shcheblykina²,

Povetka³

et al. 2021

RRP

View full text Add to dashboard Cite

show abstract

“…Plasmids permit the insertion of a DNA sequence up to 20 kb and have been used for the generation of transgenic animals. 13 The commercial pBC1 vector with 21.6 kb of DNA has been used directly or modified to generate transgenic cattle. 14 The lentiviral vectors have a restricted capacity to insert DNA, and the ideal size of genetic material to be packaged into lentiviral particles is approximately 10 kb in length.…”

Section: Vectors Used In the Generation Of Transgenic Animalsmentioning

confidence: 99%

“…29 Others studies have used MEC cultures to evaluate the expression of the vectors constructed. 13,18,19,30 Although transgenic mice may be the best model for transgenic farm animals, MEC culture is a strategy that is easier to execute than the generation of transgenic mice and shows certain advantages, such as a lower cost and an elimination of the necessity for a vivarium to produce and house transgenic mice.…”

Section: Vectors Used In the Generation Of Transgenic Animalsmentioning

confidence: 99%

Transgenic bovine as bioreactors: Challenges and perspectives

et al. 2016

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The use of recombinant proteins has increased in diverse commercial sectors. Various systems for protein production have been used for the optimization of production and functional protein expression. The mammary gland is considered to be a very interesting system for the production of recombinant proteins due to its high level of expression and its ability to perform post-translational modifications. Cows produce large quantities of milk over a long period of lactation, and therefore this species is an important candidate for recombinant protein expression in milk. However, transgenic cows are more difficult to generate due to the inefficiency of transgenic methodologies, the long periods for transgene detection, recombinant protein expression and the fact that only a single calf is obtained at the end of each pregnancy. An increase in efficiency for transgenic methodologies for cattle is a big challenge to overcome. Promising methodologies have been proposed that can help to overcome this obstacle, enabling the use of transgenic cattle as bioreactors for protein production in milk for industry.

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“…Meantime, the cell line is also used for the study of synthesis of milk fat (Harvatine, Boisclair, & Bauman, 2014;Liang et al, 2014) and milk protein (Ball, Friis, Schoenenberger, Doppler, & Groner, 1988). Furthermore, it has been used to produce recombinant proteins by transferring exogenous gene (Kaushik et al, 2014). In addition, it can be transformed into immortalized cell line by introducing immortalized gene (Chen et al, 2016;Sun et al, 2009).…”

mentioning

confidence: 99%

Isolation, characterization, and SREBP1 functional analysis of mammary epithelial cell inbuffalo

Chen

Jia

et al. 2019

J Food Biochem

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Compared to cow milk, buffalo milk contains more protein, fat, and vitamin. Buffalo milk is an ideal food in human life. Sterol regulatory element‐binding protein 1 (SREBP1), an important transcription factor, regulates the expression and activity of enzyme and protein involved in milk fat synthesis to influence on the synthesis and secretion of triglyceride in mammary epithelial cells. In the present study, we successfully isolated buffalo mammary epithelial cell by using enzymatic digestion, and then described the growth characteristics and expression characteristics of mammary epithelial cells. Moreover, we cloned the SREBP1 gene from total RNA isolated from milk fat globule and analyzed the function of the SREBP1 gene. After infected with shRNA‐SREBP1 lentiviral particle and treated with fatty acid, the expression trend of ACACA, FABP3, FAS, SCD, ERK1, ERK2, PPARy, and Insigl genes was consistent with the expression trend of SREBP1 gene. These results suggested that SREBP1 gene is a central transcription factor in regulating milk fat synthesis and SREBP1 gene may act on ERK1/ERK2 signaling pathway to regulate the expression of PPARy gene. The current study will provide a theoretical basis for further reveal the molecular mechanism of milk fat synthesis in buffalo mammary epithelial cells. Practical applications This study aim to separate and analysis characterization of mammary epithelial cell in buffalo. Compared to cow milk, buffalo milk contains more protein, fat, and vitamin. Buffalo milk is an ideal food in human life. This study will provide a theoretical basis for further research on the molecular mechanism of milk fat synthesis in buffalo mammary epithelial cells.

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Construction of a recombinant human insulin expression vector for mammary gland-specific expression in buffalo (Bubalus bubalis) mammary epithelial cell line

Cited by 7 publications

References 51 publications

Genetically modified animals for use in biopharmacology: from research to production

Genetically modified animals for use in biopharmacology: from research to production

Transgenic bovine as bioreactors: Challenges and perspectives

Isolation, characterization, and SREBP1 functional analysis of mammary epithelial cell inbuffalo

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