六方向核聚酰胺-胺树形高分子的合成及其介导的基因转移

设计、合成、表征以肌醇为六方向起始核的新型聚酰胺-胺树形高分子. 第三代至第六代树形高分子(G3~G6)能够和DNA通过电荷相互作用形成聚电解质复合物, 聚酰胺-胺树形高分子G6和荧光素酶基因(pRE Luc)形成的复合物的粒径为80~300 nm. G3~G6能介导半乳糖苷酶基因(pSV β-Gal)和荧光素酶基因有效转染HeLa和HEK293细胞. G5和G6的转染效率比聚赖氨酸高得多, 接近枝化聚乙烯亚胺(branched polyethylenimine, PEI, Mw 25 K)的转染效率. G5和G6的细胞毒性低于聚-L-赖氨酸.     

猪瘟病毒石门株E2基因的克隆及在大肠杆菌中的表达

采用ＲＴ－ＰＣＲ技术扩增了猪瘟病毒石门株的Ｅ２基因．扩增片段大小为１１８４ｂｐ，与预期大小一致．经ＳａｃＩ酶切和巢式ＰＣＲ证实了Ｅ２基因的特异性．将Ｅ２基因扩增片段首先克隆至ｐＧＥＭ－Ｔ载体中，进行全序列测定，将该基因再克隆到表达载体ｐＢＶ２２１．采用温控诱导，ＳＤＳ－ＰＡＧＥ结果显示Ｅ２基因获得表达．ＥＬＩＳＡ表明Ｅ２基因表达产物可与猪瘟阳性血清起特异性反应．     

伪狂犬病毒表达载体的构建

利用伪狂犬病毒（Ｐｓｅｕｄｏｒａｂｉｅｓｖｉｒｕｓ，ＰＲＶ）湖北地方株胸腺核苷激酶（ＴＫ）基因和ｇＧ糖蛋白基因启动子（ＰｇＧ），构建了ＰＲＶ基因转移载体．通过β－半乳糖苷酶基因确定ＰｇＧ控制下外源基因的表达并筛选出表达β－半乳糖苷酶的ＰＲＶ重组病毒．采用ＰＣＲ对重组病毒进行了初步鉴定，证实外源基因可由构建的转移载体导入病毒ｔｋ基因中，重组的ＰＲＶ湖北地方株可用于表达外源基因．     

Syntheses of polyamidoamine dendrimers starting from a hexadimensional core and application in gene transfer

Gene therapy refers to the concept and practice of applying gene to treat diseases. It may be defined as a method for inserting a functioning gene into the cells of a patient to correct an inborn error of metabolism (i.e. genetic abnormality or birth defect) or to provide a new function in a cell. There are numerous diseases that may be treated by gene therapy including genetic defects, com-mon illnesses such as cancer, AIDS and chronic diseases such as diabetes[1]. A gene medicine sys-tem c…