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1

Structure, expression and engineering of milk protein genes

100%
Zwierzchowski L.
Biotechnologia
|
1996
|
issue 2
81-94
EN
Expression of milk protein genes is regulated by hormones, growth factors and extracellular matrix.Prolactin, the major lactogenic hormone, promotes all stages of casein gene expression. Besides prolactin, also growth hormone may directly induce expression of milk protein genes. Computer analysis, mutation experiments and DNA-protein binding experiments enabled indentification of mammary-specific trasnscription factors and cis-regulatory sequences in milk protein gene promoters. Also, transgene technology enables recognition of regulatory sequences in milk protein genes.Moreover, transgenic animals carrying structural genes of human proteins fused to mammary specific promoters are considered as living "bioreactors" designated to produce human proteins for pharmaceutical use.
2

MilkProtChip ? a microarray of SNPs in candidate genes associated with milk protein biosynthesis ? development and validation

80%
Kaminski S., Ahman A., Rusc A., Wojcik E., Malewski T.
Journal of Applied Genetics
|
2005
|
vol. 46
|
issue 1
45-58
EN
MilkProtChip is an oligonucleotide microarray based on the arrayed primer extension (APEX) technique, allowing genotyping of single nucleotide polymorphisms (SNPs) in genes of interest for bovine milk protein biosynthesis. APEX consists of a sequencing reaction primed by an oligonucleotide anchored with its 5'end to a glass slide and terminating one nucleotide before the polymorphic site. The extension with one fluorescently labeled dideoxy nucleotide complementary to the template reveals the polymorphism. A total of 75 SNPs were selected among those associated directly or potentially with milk protein content. Among the 75 SNPs, 4 did not produce a positive signal. Most of the remaining SNPs produced a signal for both strands, except for 4 (one strand). In the validation step, 12 Polish Holstein bulls, 1 Polish Red bull, 1 bison (Bison bonasus), 11 Jersey cows and 25 Polish Holstein cows were screened to validate SNPs. Among the 71 selected SNPs ? 26 were found monoallelic, the rest showing at least two genotypes for the entire population under study. All the animals were earlier genotyped for 2?5 SNPs by PCR-RFLP and PCR sequencing and all showed complete concordance with APEX genotyping. APEX reactions showed relatively high signal frequencies: more than 0.9, 0.9?0.8 and below 0.8, for 65, 4 and 2 DNA samples, respectively. The primary application of the MilkProtChip is the simultaneous genotyping of dozens of SNPs to reveal and clarify the genetic background of milk protein biosynthesis. The chip may possibly be used for dairy cattle identification and paternity analysis, evolutionary studies, the evaluation of genetic distances between wild and domestic cattle breeds and the domestication history of bovine species.
3

Biotechnology of the mammary gland - perspectives of genetic engineering of milk proteins

80%
Zwierzchowski L.
Biotechnologia
|
1998
|
issue 2
33-56
EN
In this article, state of art and perspectives in mammary gland biotechnology are reviewed. Recent progress in recombinant DNA technology as well in embryo manipulation and transfer has made the introduction of specific genes into the germline of animals relatively easy. With appropriate genetic constructs, the expression of the inserted genes in transgenic animals can be controlled in a tissue-specific and in a differentiation-specific manner. Thus, it is now possible to consider alteration of the composition of milk produced by lactating animals in a variety of ways. There is a growing list of foreign milk proteins that have been expressed, and one can envisage placing almost any protein gene of interest under the control of promoter of a milk protein gene. Many human proteins of a potential pharmaceutical use may be now produced in the mammary glands of laboratory or farm animals. Modification of milk composition can be extended not only to produce proteins of commercial value but also, by manipulation of key metabolic enzymes, to fat, lactose, and other components of milk. Many alternations in ruminants' milk composition, including 'humanization' of cow's milk, are planned, however, these manipulations must avait the development of totipotent embryonic cell lines (ESC) of farm animals, cells that enable gene manipulation by homologous recombination. In spite of a great progress, many obstacles and difficulties still exist on the way to economical production of human paharmaceuticals in farm 'transgenic bioreactoractors'. These dificulties are discussed in detail.
4

Associations between bovine beta-lactoglobulin polymorphism within coding and regulatory sequences and milk performance traits

80%
Kaminski S., Zabolewicz T.
Journal of Applied Genetics
|
2000
|
vol. 41
|
issue 2
91-99
EN
The bovine beta-lactoglobulin (LGB) gene is considered a potential quantitative trait locus in dairy cattle breeding. In Black-and-White dairy cattle the LGB gene has two predominant alleles A and B. This can result in three possible genotypes AA, AB and BB. Moreover, within the promoter of the gene several point mutations were found. A herd of one hundred and twenty-four Black-and-White cows were genotyped for two loci: locus LGB (exon IV, alleles A and B) and locus LGB-R (SSCP polymorphism within a fragment of LGB promoter: SSCP patterns R2, R3, R1, R9). In our sample 13 AA, 58 AB and 53 BB LGB cows and 66 R2, 16 R3, 40 R1 and 2 R9 LGB-R cows were identified. A statistical analysis revealed significant associations between LGB, LGB-R genotypes as well as intragenic haplotypes LGB/LGB-R and milk protein content during the first complete lactation. Cows with AA LGB genotype, R3 LGB-R SSCP pattern and AA/R3 haplotypes had the highest protein content. These results support the hypothesis that sequence variation within the promoter of the LGB gene is probably one of the factors responsible for differences in milk protein content.
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