کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10881851 | 1077926 | 2005 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Gene transfection and expression in a primary culture of mammary epithelial cells isolated from lactating sows
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کلمات کلیدی
eGFPRT-PCRβ-LgBMECsα-La - α-لاα-Lactalbumin - α-لاکتبالومینβ-Lactoglobulin - β-لاکتوگلوبولینβ-Casein - β-کازئینGene transfection - انتقال ژنGene expression - بیان ژنPig - خوکMammary epithelial cells - سلول های اپیتلیال پستانBovine mammary epithelial cells - سلول های اپیتلیال پستان مولد گاوreverse transcription-polymerase chain reaction - واکنش زنجیره ای رونویسی-پلیمراز معکوسenhanced green fluorescent protein - پروتئین فلورسنت سبز افزایش یافته است
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوفیزیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Porcine mammary epithelial cells (PMECs) were isolated from lactating sow mammary glands and cultured on a matrix gel. Primary culture cells expressed significant amounts of the specific marker cytokeratin as determined by immunohistochemistry, and exhibited mammary-specific functions, such as transcription of α-lactalbumin, β-casein and β-lactoglobulin genes. They also formed mammospheres when the medium was supplemented with lactogenic hormones. The PMECs were used to study gene transfer and expression in vitro. A gene encoding enhanced green fluorescent protein (EGFP) was used as a reporter and two constructs were investigated, pEGFP-N1 (a vector constructed with a CMV promoter followed by the EGFP gene) and pGB562/GFP (a mammary gland-specific expression vector with regulatory sequences from the goat β-casein gene linked to EGFP). The efficiency of DNA transfer into the cultured PMECs was about 20-30%. GFP expression in the pGB562/GFP-transfected PMECs was markedly stimulated by prolactin supplements in the medium. The established PMECs maintained optimal gene expression from 1 to 20 passages and appeared to provide an efficient and convenient system for assessing the expression of transgenes containing mammary gland-specific promoters.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Cell Biology International - Volume 29, Issue 7, July 2005, Pages 576-582
Journal: Cell Biology International - Volume 29, Issue 7, July 2005, Pages 576-582
نویسندگان
Y.L. Sun, C.S. Lin, Y.C. Chou,