Preparation of recombinant proteins in milk to improve human and animal health

Soler, Éric; Thépot, Dominique; Rival‐Gervier, Sylvie; Jolivet, Geneviève; Houdebiné, Louis‐Marie

doi:10.1051/rnd:2006029

Cited by 20 publications

(20 citation statements)

References 43 publications

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“…Similar tools may also be used to improve animal production, including the modification of milk composition for human and animal consumption (Bleck et al, 1998;Hiripi et al, 2000;Brophy et al, 2003;Houdebine, 2004;Soler et al, 2006;Baranyi et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

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Rabbit milk protein genes: from mRNA identification to chromatin structure

Jolivet

Daniel-Carlier

Thépot

et al. 2008

Animal

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Milk protein genes are among the most intensively expressed and they are active only in epithelial mammary cells of lactating animals. They code for proteins which represent 30% of the proteins consumed by humans in developed countries. Mammary gland development occurs essentially during each pregnancy. This offers experimenters attractive models to study the expression mechanisms of genes controlled by known hormones and factors (prolactin, glucocorticoids, progesterone, insulin-like growth factor-1 and others) as well as extracellular matrix. In the mid-1970s, it became possible to identify and quantify mRNAs from higher living organisms using translation in reticulocyte lysate. A few years later, the use of radioactive cDNAs as probes made it possible for the quantification of mRNA in various physiological situations using hybridisation in the liquid phase. Gene cloning offered additional tools to measure milk protein mRNAs and also to identify transcription factors. Gene transfer in cultured mammary cells and in animals contributed greatly to these studies. It is now well established that most if not all genes of higher eukaryotes are under the control of multiple distal regulatory elements and that local modifications of the chromatin structure play an essential role in the mechanisms of differentiation from embryos to adults. The technique, known as ChIP (chromatin immunoprecipitation), is being implemented to identify the factors that modify chromatin structure at the milk protein gene level during embryo development, mammogenesis and lactogenesis, including the action of hormones and extracellular matrix. Transgenesis is not just a tool to study gene regulation and function, it is also currently used for various biotechnological applications including the preparation of pharmaceutical proteins in milk. This implies the design of efficient vectors capable of directing the secretion of recombinant proteins in milk at a high concentration. Milk protein gene promoters and long genomic-DNA fragments containing essentially all the regulatory elements of milk protein genes are used to optimise recombinant protein production in milk.Keywords: chromatin, development, promoter, transgenesis IntroductionMilk protein genes are abundantly and specifically expressed in the mammary gland of lactating mammals. Lactation is essential for the development of newborns. Since the domestication of a few ruminants, milk has become an important food for humans. Milk proteins represent 30% of the proteins consumed by humans in developed countries. On the other hand, milk protein genes are among the most intensively expressed genes and they were soon candidates to study the mechanisms of gene expression. For these reasons, biology of lactation has been extensively studied for the last 50 years. The main factors controlling mammary gland development and milk protein gene expression are well known. These factors are oestrogens, prolactin, progesterone, glucocorticoids, insulin-like growth factor-1, placental lactogen and extracellu...

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Section: Discussionmentioning

confidence: 99%

“…The EC SOD was not fully sialylated but highly active, whereas the porcine LH/CG was active in vitro but not in vivo due to its incomplete glycosylation and short half-life. Such observations have been conducted more recently for other proteins produced in rabbit milk (Koles et al, 2004;Soler et al, 2006;Houdebine, 2007a).…”

Section: Use Of Transgenesismentioning

confidence: 93%

Rabbit milk protein genes: from mRNA identification to chromatin structure

Jolivet

Daniel-Carlier

Thépot

et al. 2008

Animal

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“…Such observation was expected considering the lactating stage of mammary gland sample tissues [22]. Functional profile also revealed the presence of genes coding for various metabolic and immune response proteins, which could be justified for their potential roles in milk digestion, nutrients absorption, and immunity development in baby animals [1,2,4,11,13,14]. Milk immune response in human, for instance, has been proven to provide passive protection to the young [23].…”

Section: Characterization Of Primary Mammary Gland Cdna Librarymentioning

confidence: 98%

“…With current genomic advances such limitations could be corrected through gene manipulations [13][14][15]. However, because of the insufficiency of genomic resources and information pertaining to goat, it is very difficult to improve this species, in general, or solve the repulsive goaty smell problem, in particular, using modern technologies.…”

Section: Introductionmentioning

confidence: 99%

Construction and Characterization of a Goat Mammary Gland cDNA Library

Han

Luo

et al. 2007

Mol Biotechnol

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Sensitive, specific, and accurate methods to assay chemosensitivity are needed to (1) screen new therapeutic agents, (2) identify patterns of chemosensitivity for different tumor types, (3) establish patterns of cross-resistance and sensitivity in treatment of naïve and relapsing tumors, (4) identify genomic and proteomic profiles associated with sensitivity, (5) correlate in vitro response with preclinical in vivo effects and clinical outcomes for a particular therapeutic agent, and (6) tailor chemotherapy regimens to individual patients. Various methods are available to achieve these end points, including several in vitro clonogenic and proliferation assays, cell metabolic activity assays, molecular assays to monitor expression of markers for responsiveness, drug resistance, and for induction of apoptosis, in vivo tumor growth and survival assays in metastatic and orthotopic models, and in vivo imaging assays. The advantages and disadvantages of the specific assays are discussed. A summary of research questions related to chemosensitivity testing is also included.

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“…It is also a possible source of pharmaceutical proteins. Yet approximately 100 recombinant proteins have been expressed in mouse milk using transgenic technology (Soler et al, 2006). So in this study, a lysine rich gene which is derived from the fragments of -casein, S2-casein and siderophilin were expressed in eukaryotic cells and cow milk to determine the feasibility of producing lysineenriched milk in transgenic cows.…”

Section: Introductionmentioning

confidence: 99%

Expression and characterization of a lysine rich protein in cow milk

Zhang

et al. 2018

IJAR

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The lysine is considered as the most important essential amino acid, because it is the most limiting in the cereals grains. In this study, a lysine-rich (LR) gene, and the expression vector pcDNA3.1-LR and pBC1-LR were constructed. The LR was expressed in 293T cells driven by the vector pcDNA3.1-LR and checked by RT-PCR and WB. The mammary gland tissuespecific expression vector carrying the LR was injected directly into the lactating mammary glands of cows and the milk samples were checked by a complete amino acid analysis. The results showed that the LR protein was expressed successfully in cells and in cow milk; the expression of LR lasted for 6 d, and the lysine level of the injection group was significantly higher than that of negative controls (p <0.05). This study provide a better understanding of how mammary gland expression systems increase the lysine content of milk that can be applied to transgenic dairy cow.

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Preparation of recombinant proteins in milk to improve human and animal health

Cited by 20 publications

References 43 publications

Rabbit milk protein genes: from mRNA identification to chromatin structure

Rabbit milk protein genes: from mRNA identification to chromatin structure

Construction and Characterization of a Goat Mammary Gland cDNA Library

Expression and characterization of a lysine rich protein in cow milk

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