Phenotypic features of first-generation transgenic goats for human granulocyte-colony stimulation factor production in milk

Batista, Ríbrio Ivan Tavares Pereira; Melo, Cynthia de Freitas; Souza-Fabjan, J. M. G.; Teixeira, Dárcio Ítalo Alves; Melo, Luciana Magalhães; Freitas, V. J. F.

doi:10.1007/s10529-014-1588-0

Cited by 8 publications

(8 citation statements)

References 16 publications

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“…On the whole, the results pointed out to the complete absence of inflammations or infections of goat mammary glands. The milk composition was similar to that reported in an earlier experiment performed by our group with Caninde goats (5) . In addition, it is important to highlight that analysis of physicochemical properties and ultrasonographic examinations of mammary glands were relatively constant for all females throughout the lactation period.…”

Section: Resultssupporting

confidence: 87%

“…Furthermore, the lactation initiated in a similar period that has alrady been cited for goats (3) . The values of average daily milk production obtained on the first thirty days of lactation were higher than those found by Batista et al (5) , who also studied milk hormonally induced Caninde goats in the same lactation period.…”

Section: Resultscontrasting

confidence: 77%

“…The induction of lactation through hormonal stimulation has been previously demonstrated in goats (3)(4)(5) .…”

Section: Introductionmentioning

confidence: 93%

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Mammary Biopsy in Induced Lactating Goats: Assessment of Post-Biopsy Milk Properties and Ultrasonographic Appearance

et al. 2017

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The aim of this study was to evaluate the effects of mammary biopsy procedures on gland function of goats submitted to hormonal lactation. Ten female Caninde goats, at the age of around 3 years, were used, and lactation was induced by using estrogen, progesterone, and prednisolone. Ultrasonographic examinations of mammary gland, milking, and analysis of physicochemical properties of milk were performed immediately before (0 h) and 24, 48, and 72 h after each biopsy procedure. Surgical mammary gland biopsies were obtained at days 5 and 26 of the lactation. Even after drying initiation, all animals produced milk for five months. No abnormalities were observed on the ultrasonographic appearance of biopsied glands. All 20 biopsies were technically successful and any inflammation or infection was associated with biopsy procedures. Regarding the physicochemical properties and volumes of milk produced, no significant differences (P > 0.05) were observed between right and left glands nor in the comparison of the results from time 0 with 24, 48, and 72 h for biopsied teats. The mammary biopsy did not induce neither severe changes in production and physicochemical properties of the milk nor in ultrasonographic appearance of mammary gland of goats in hormonal lactation.Keywords: biopsy; goat; udder; ultrasonography. ResumoO objetivo deste estudo foi avaliar os efeitos dos procedimentos de biópsia mamária sobre a função da glândula de caprinos submetidos à lactação induzida. Dez cabras Canindé com idade de cerca de 3 anos foram utilizadas e a lactação foi induzida pelo uso de estrógeno, progesterona e prednisolona. Exames ultrassonográficos da glândula mamária, ordenha e análise das propriedades físico-químicas do leite foram realizados imediatamente antes (0 h) e 24, 48 e 72 h após cada procedimento de biópsia. As biópsias cirúrgicas de glândula mamária foram obtidas aos dias 5 e 26 da lactação. Mesmo após o início do processo de secagem, todos os animais continuaram produzindo leite por cinco meses. Nenhuma anormalidade foi observada na aparência ultrassonográfica das glândulas biopsiadas. Todas as 20 biópsias foram realizadas com sucesso e nenhuma inflamação ou infecção foi observada. Com relação às propriedades físico-químicas e volumes de leite produzido, nenhuma diferença significativa Cienc. anim. bras., Goiânia, v.18, 1-7, e-39077, 2017 2 SOUSA, F.C. et al.(P > 0,05) foi observada entre a glândula direita e esquerda, nem comparando resultados do momento 0 com 24, 48 e 72 h para as glândulas biopsiadas. A biópsia mamária não induz severas mudanças na produção e nas propriedades físico-químicas do leite nem na aparência ultrassonográfica da glândula mamária de caprinos em lactação.

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Section: Resultssupporting

confidence: 87%

Section: Resultscontrasting

confidence: 77%

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Mammary Biopsy in Induced Lactating Goats: Assessment of Post-Biopsy Milk Properties and Ultrasonographic Appearance

et al. 2017

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“…Owing to the high expression level, low cost, ease of purification[ 6 ], ideal post-translation modification similar to that of human proteins, and higher biological activity, the mammary gland bioreactor overcomes nearly all the shortcomings and disadvantages of other expression systems. Therefore, medicinal recombinant proteins expressed in transgenic animal mammary gland bioreactors are gaining the attention of researchers and have wide development prospects[ 14 – 17 ] . Animal mammary glands have become the most ideal expression systems to produce recombinant proteins.…”

Section: Introductionmentioning

confidence: 99%

A novel recombinant human plasminogen activator: Efficient expression and hereditary stability in transgenic goats and in vitro thrombolytic bioactivity in the milk of transgenic goats

Lü

Zhang

et al. 2018

PLoS ONE

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BackgroundThromboses is a rapidly growing medical problem worldwide. Low-cost, high-scale production of thrombotic drugs is needed to meet the demand. The production of biomolecules in transgenic animals might help address this issue. To our knowledge, the expression of recombinant human plasminogen activator (rhPA) in goat mammary glands has never been reported before.MethodsWe constructed a mammary gland–specific expression vector, BLC14/rhPA, which encodes only the essential K2 fibrin-binding and P domains of wild-type tPA (deletion mutant of tPA lacking the F, E, and K1 domains), along with the goat β-lactoglobulin gene signal peptide-coding sequence. The mammary gland–specific expression vector BLC14/rhPA was transfected into goat fetal fibroblast cells by electroporation. After selection for 3 weeks by G418, stably transfected cell colonies were obtained. PCR analysis results indicated that 24 of the resistant clones were transgenic cell lines; of these, 8 lines were selected as the donor cells. The positive cells were starved for 72 h with DMEM/F12 medium containing 0.5% FBS and were then used as do. Finally, 256 reconstructed oocytes were transferred into 26 recipients, and 7 of them became pregnant (pregnancy rate, 26.9%). Two kids were obtained (BP21 and BP22). PCR analysis confirmed that both were transgenic goats. To analyze the heredity of the rhPA expressed in BP21 F0 and F1 transgenic goats, the F0 transgenic goat BP21 was mated with a normal male goat to generate an F1 transgenic goat. Enucleated metaphase II (MII) oocytes and positive donor cells were used to reconstruct embryos, which were transplanted into the oviducts of the recipients.ResultsWestern blot results showed a specific 39 kDa band. The rhPA expression level in transgenic goat whey was about 78.32 μg/mL by ELISA. Results of ELISA and the in vitro thrombolysis test (FAPA) showed that specific activity of the rhPA in the milk of F0 and F1 transgenic goats was 13.3 times higher than that of the reteplase reference material.ConclusionThus, we demonstrated that BLC14/rhPA was reasonably effective for expression in the mammary glands of transgenic goats, and was stably inherited by the offspring. This study provides the basis for the large-scale production of biological pharmaceuticals in transgenic animals. The expression of biopharmaceuticals by transgenic animals can be used for pharmacological research and bioactive analysis, and transgenic goats were demonstrated to be promising animals for the large-scale production of thrombolytic biopharmaceuticals.

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“…It is of note that many active human proteins for medical applications are produced in the milk of transgenic animals (Li et al 2013;Batista et al 2014;Park et al 2015).…”

Section: Introductionmentioning

confidence: 99%

The development of modified human Hsp70 (HSPA1A) and its production in the milk of transgenic mice

Gurskiy

Гарбуз

Сошникова

et al. 2016

Cell Stress and Chaperones

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The production of major human heat shock protein Hsp70 (HSPA1A) in a eukaryotic expression system is needed for testing and possible medical applications. In this study, transgenic mice were produced containing wild-type human Hsp70 allele in the vector providing expression in the milk. The results indicated that human Hsp70 was readily expressed in the transgenic animals but did not apparently preserve its intact structure and, hence, it was not possible to purify the protein using conventional isolation techniques. It was suggested that the protein underwent glycosylation in the process of expression, and this quite common modification for proteins expressed in the milk complicated its isolation. To check this possibility, we mutated all presumptive sites of glycosylation and tested the properties of the resulting modified Hsp70 expressed in E. coli. The investigation demonstrated that the modified protein exhibited all beneficial properties of the wild-type Hsp70 and was even superior to the latter for a few parameters. Based on these results, a transgenic mouse strain was obtained which expressed the modified Hsp70 in milk and which was easy to isolate using ATP columns. Therefore, the developed construct can be explored in various bioreactors for reliable manufacture of high quality, uniform, and reproducible human Hsp70 for possible medical applications including neurodegenerative diseases and cancer.

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Phenotypic features of first-generation transgenic goats for human granulocyte-colony stimulation factor production in milk

Cited by 8 publications

References 16 publications

Mammary Biopsy in Induced Lactating Goats: Assessment of Post-Biopsy Milk Properties and Ultrasonographic Appearance

Mammary Biopsy in Induced Lactating Goats: Assessment of Post-Biopsy Milk Properties and Ultrasonographic Appearance

A novel recombinant human plasminogen activator: Efficient expression and hereditary stability in transgenic goats and in vitro thrombolytic bioactivity in the milk of transgenic goats

The development of modified human Hsp70 (HSPA1A) and its production in the milk of transgenic mice

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