泉州师范学院学报
泉州師範學院學報
천주사범학원학보
JOURNAL OF QUANZHOU NORMAL COLLGEG
2011年
4期
45-49
,共5页
铁蛋白%重组%表达
鐵蛋白%重組%錶達
철단백%중조%표체
ferritin%reconstruction%expression
通过PCR技术扩增出人铁蛋白基因,经酶切后与表达载体质粒pGEX-4T-2连接,重组质粒转化感受态大肠杆菌,利用菌落PCR、质粒双酶切、测序,证实成功地构建了人铁蛋白基因表达载体,利用IPTG对重组菌进行诱导表达,通过尿素洗涤纯化目的蛋白用于制备抗体.
通過PCR技術擴增齣人鐵蛋白基因,經酶切後與錶達載體質粒pGEX-4T-2連接,重組質粒轉化感受態大腸桿菌,利用菌落PCR、質粒雙酶切、測序,證實成功地構建瞭人鐵蛋白基因錶達載體,利用IPTG對重組菌進行誘導錶達,通過尿素洗滌純化目的蛋白用于製備抗體.
통과PCR기술확증출인철단백기인,경매절후여표체재체질립pGEX-4T-2련접,중조질립전화감수태대장간균,이용균락PCR、질립쌍매절、측서,증실성공지구건료인철단백기인표체재체,이용IPTG대중조균진행유도표체,통과뇨소세조순화목적단백용우제비항체.
The ferritin gene was cloned from human genome using the PCR technology. After digested by the enzymes (BamH I, Sma I), it was inserted into the plasmid pGEX-4T-2 to reconstruct the expression vector. Then the reconstructed plasmid was converted into E. coli whereby to confirm using the colony PCR, the enzyme digestion and the DNA sequencing. The recombinant protein was expressed after inducing by IPTG and then purified using urea for antibody preparation.