中文摘要：

目的：克隆人抗酶抑制因子-1（omithine decarboxylase antizyme inhibitor—1，OAZI-1）cDNA，建立在大肠杆菌中原核表达并纯化人OAZI-1蛋白的实验技术。方法：巢式RT—PCR法从人A549总RNA中扩增人OAZI—1 cDNA并构建pET-28a／OAZI-1原核表达质粒。该质粒转化大肠杆菌原核表达菌BL21（DE3）后IPTG诱导表达。诱导表达出的重组蛋白用Ni—NTA树脂亲和层析纯化。SDS—PAGE和Western法检测重组OAZI—1蛋白的表达和纯化。结果：成功克隆出编码全长人OAZI-1的eDNA序列，并构建出原核表达质粒pET-28a／OAZI—1。DNA测序分析，重组质粒中的OAZI—1 cDNA无突变，与6XHis标签框架对接正确。重组质粒转化人大肠杆菌表达菌BL21（DE3）中后，可用IPTG诱导表达出重组OAZI—1蛋白，该重组蛋白可用Ni—NTA树脂亲和层析纯化。结论：成功建立了人抗酶抑制因子的原核表达和纯化的实验方法，为后续OAZI-1的功能研究奠定了基础。

英文摘要：

Objective： To clone the human ornithine antizyme intfibitor - 1 （OAZI - 1 ） cDNA and establish the method for prokaryotic expression and purification of the OAZI - 1. Method ： OAZI - 1 cDNA was cloned out from total RNA of human A549 lung cancer line by nest RT- PCR and then subcloned into the prokaryotic expression vector pET -28a （ ＋ ）. Resulted plasmid pET -28a/OAZI - 1 was transformed into E. coli. BL21 （DE3）. Expression of OAZI - 1 in the host cells was induced by IPTG. The recombinant OAZI - 1 was purified by Ni - NTA and identified by SDS - PAGE and western blot assay. Result： The OAZI - 1 cDNA was successfully obtained and cloned into the vector pET- 28a（ ＋ ）. DNA sequencing proved that there were no mutations and frameshift for OAZI - 1 insert in pET- 28a/OAZI - 1. When the plasmid was transformed into E. coil, the recombinant OAZI - 1 could be expressed in the host cells by IPTG induction and purified by Ni - NTA resin. Conclusion ： A method for prokaryotic expression and purification of human OAZI - 1 was successfully established, whicb lays a foundation for the OAZI - 1 functional research.