The goat ßcasein/CMV chimeric promoter drives the expression of hLF in transgenic goats produced by cell transgene microinjection.
Int J Mol Med
; 44(6): 2057-2064, 2019 Dec.
Article
em En
| MEDLINE
| ID: mdl-31661123
ABSTRACT
There is growing interest in the application of lactoferrin (LF) as a drug or food additive for animals and humans. The objective of this study was to produce transgenic cloned goats that would serve as living bioreactors, expressing high levels of recombinant human LF (rhLF) in their milk. We designed a pCL25 expression vector containing goat ßcasein/CMV chimeric promoter in order to facilitate rhLF expression. This pCL25rhLFNeo vector was microinjected into goat fetal fibroblasts. G418 selection and PCR analysis were used to identify transgenic donor cells suitable for somatic cell nuclear transfer (SCNT). After SCNT and embryo transplantation, goats harboring the hLF gene were produced, as confirmed via PCR and southern blotting. The average rhLF concentration in milk from this transgenic goat was 3.89 mg/ml as determined via ELISA. We also used an optimized buffer in order to effectively elute highpurity (95.8%) rhLF from a cationexchange column, with the recovered rhLF exhibiting high biological activity. Findings from this study demonstrated that it is possible to generate a transgenic goat harboring the hLF transgene driven by the goat ßcasein/CMV chimeric promoter. It represents an initial step towards the production of rhLF, potentially allowing for industrialized purification in the future.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Animais Geneticamente Modificados
/
Regiões Promotoras Genéticas
/
Transgenes
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Lactoferrina
Tipo de estudo:
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article