人血清白蛋白DNA片段的扩增及克隆

Polymerase Chain Reaction and Clone of Human Serum Albumin Gene Fragments

根据人血清白蛋白(HSA)基因的第八个外显子区DNA序列及相应cDNA序列和KB小鼠血清蛋白DNA序列设计的一对引物P_1:5'-GGATCCACCAACTTACTTATAGGCG-3'和P_2:5'-AGGATCCTACTTACATGCCCAGGAAG-3'.以人白细胞和KB鼠DNA为模板,在50_(μl)PCR反应体系中,经94℃、lmin,58.5℃,35S,72℃、lmin三个循环.94℃、lmin,57.5℃,40S.72℃、lmin32个循环扩增,获得了约为256bp的单一带DNA,KB鼠无专一带.将人HSA DNA扩增的单一带DNA以BamHI-Sst I位点克隆到pUC19中,得到HSA DNA扩增片段的克隆子.

Designing a pair of primers, P1:5' -GGATCCACCAACTTACTTATAGGCG-3' and P2:5' - AGGATCCTACTTACATGCCCAGGAAG-3' according to Human and mousegenome. Template DNA is from human white cell and KB mouse DNA with phenol purified. Polymerase chain reaction (PCR) system is volume 50ul, 94℃ denaturation for 1min,58℃annealing 35 s, 72℃ extension 1min, 35 cycles. about 256 bp band was obtained, KB mouse has not the result. 256 bp DNA was ligased with pud19 got Clone.