内蒙古大学学报(自然科学版)
內矇古大學學報(自然科學版)
내몽고대학학보(자연과학판)
JOURNAL OF INNER MONGOLIA AGRICULTURAL UNIVERSITY
2001年
1期
79-82
,共4页
李雪玲%郭继彤%于海泉%王达珍%张肇英
李雪玲%郭繼彤%于海泉%王達珍%張肇英
리설령%곽계동%우해천%왕체진%장조영
绵羊β-乳球蛋白基因(BLG)5′端及上游调控序列%PCR
綿羊β-乳毬蛋白基因(BLG)5′耑及上遊調控序列%PCR
면양β-유구단백기인(BLG)5′단급상유조공서렬%PCR
本文对绵羊β-乳球蛋白基因5′端及上游调控序列的PCR方法扩增进行了研究.经比较分析，将从羊血中提取的绵羊基因组DNA的模板用量定为4μl(0.4μg/μl),TaqDNA聚合酶用量为0.83μl(3u/μl),变性为94℃1min，退火为64℃1min，72℃延伸2min，循环次数为32次，获得的PCR产物经电泳检测，条带明亮，特异性高，大小为898bp.经序列分析发现，与已知基因组序列一致性达99%以上，可用于指导外源基因在转基因动物乳腺中表达.
本文對綿羊β-乳毬蛋白基因5′耑及上遊調控序列的PCR方法擴增進行瞭研究.經比較分析，將從羊血中提取的綿羊基因組DNA的模闆用量定為4μl(0.4μg/μl),TaqDNA聚閤酶用量為0.83μl(3u/μl),變性為94℃1min，退火為64℃1min，72℃延伸2min，循環次數為32次，穫得的PCR產物經電泳檢測，條帶明亮，特異性高，大小為898bp.經序列分析髮現，與已知基因組序列一緻性達99%以上，可用于指導外源基因在轉基因動物乳腺中錶達.
본문대면양β-유구단백기인5′단급상유조공서렬적PCR방법확증진행료연구.경비교분석，장종양혈중제취적면양기인조DNA적모판용량정위4μl(0.4μg/μl),TaqDNA취합매용량위0.83μl(3u/μl),변성위94℃1min，퇴화위64℃1min，72℃연신2min，순배차수위32차，획득적PCR산물경전영검측，조대명량，특이성고，대소위898bp.경서렬분석발현，여이지기인조서렬일치성체99%이상，가용우지도외원기인재전기인동물유선중표체.
This study was carried on the amplification of BLG 5′-flankingand regulatory sequence by PCR. The template was the sheep genomic DNA extracted from blood of sheep. The denaturation temperature was 94 ℃ 1 min,the annealling temperature was 64℃ 1min,the extention temperature was 72 ℃ 2 mins. After 32 cycles,the product was analyzed by agarose gel electrophoresis,and sequence assay ,and extracted and used to construct new gene with hCD14. The results shows that this PCR product shares 99% nucleotide sequence with the reported nucleotide sequence of the genomic ovine BLG and can direct the mammary specific expression of hCD14 in transgenic mice.
