蓖麻毒素A链的基因克隆  

Cloning of the Ricin A Chain Gene

作　　者：王文香[1] 汲言山[1] 沈倍奋[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《生物化学杂志》1993年第5期518-521,共4页

摘　　要：作为导向药物蓖麻毒素A(Ricin-A)链在大肠杆菌中表达时不含糖基侧链,在体内半衰期长,可提高共作为导向药物的疗效。我们根据Ricin基因核苷酸序列,设计Ricin-A的上、下游引物,通过PCR(多聚酶链式反应)方法,扩增出Ricin-A链基因。与pUC_(19)载体连接,转化到JM103大肠杆菌中,得到重组克隆。对其进行几种酶切鉴定,证明酶切位点正确,又经序列分析,读出与文献发表的Ricin-A序列只有两个碱基不同,但无氨基酸残基的改变。有关Ricin-A的表达工作正在进行中。Ricin is the ribosome-inactivating protein in Ricinus communis. The rapid clearance of the native ricin-A chain is primarily due to the recognition of mannose or fucose residues on the A chain component by cells with mannose or fucose receptors in liver and other organs. An effective solution to the problem is to use recombinant ricin A chains expressed in bacteria. In the present paper, we synthesized the primers for polymerase chain reaction, according to the sequence of the ricin A chain. The gene obtained by PCR was cloned into pUC_(19) plasmid and sequenced by T7 polymerase sequencing kit. The resulted nucleotide seqeunce is the same as that of ricin A, except two nucleotides.

关 键 词：基因克隆 蓖麻毒素 蓖麻毒素A 

分 类 号：Q7[生物学—分子生物学]