Department of Biochemistry and Biotechnology, Poznan University of Life Sciences, Dojazd 11, 60-632, Poznan, Poland.
J Appl Genet. 2012 Nov;53(4):435-42. doi: 10.1007/s13353-012-0110-4. Epub 2012 Aug 17.
The advent of transgenic technology has provided methods for the production of pharmaceuticals by the isolation of these proteins from transgenic animals. The mammary gland has been focused on as a bioreactor, since milk is easily collected from lactating animals and protein production can be expressed at very high levels, including hormones and enzymes. We demonstrate here the expression pattern of recombinant human growth hormone (rhGH) in transgenic rabbits carrying hGH genomic sequences driven by the rat whey acidic protein (WAP) promoter. The transgene was mapped to the q26-27 telomere region of chromosome 7q by fluorescence in situ hybridization (FISH). Nearly 30 % of the F1 generation demonstrated the presence of transgene. The recombinant growth hormone was detected in the milk of the transgenic rabbit females, but not in serum, up to the level of 10 μg/ml. Ectopic expression of the transgene in the brain, heart, kidney, liver, and salivary gland was not observed, indicating that a short sequence of rat WAP promoter (969 bp) contained essential sequences directing expression exclusively to the mammary gland. The biological activity of recombinant growth hormone was measured by immunoreactivity and the capability to stimulate growth of the hormone-dependent Nb211 cell line.
转基因技术的出现为从转基因动物中分离这些蛋白质来生产药物提供了方法。由于从泌乳动物中很容易收集乳汁,并且可以非常高的水平(包括激素和酶)表达蛋白质生产,因此乳腺已成为生物反应器的焦点。我们在这里展示了携带由大鼠乳清酸性蛋白(WAP)启动子驱动的人重组生长激素(rhGH)基因组序列的转基因兔中重组人生长激素(rhGH)的表达模式。通过荧光原位杂交(FISH)将转基因定位到染色体 7q 的 q26-27 端粒区。将近 30%的 F1 代显示出存在转基因。在转基因兔的雌性乳汁中检测到重组生长激素,但在血清中未检测到,其水平高达 10μg/ml。未观察到转基因在脑,心脏,肾脏,肝脏和唾液腺中的异位表达,这表明大鼠 WAP 启动子(969bp)的短序列包含指导乳腺中表达的必需序列。通过免疫反应性和刺激激素依赖性 Nb211 细胞系生长的能力来测量重组生长激素的生物学活性。