重组人白细胞介素-3真核表达质粒直接注射小鼠骨骼肌的体内表达研究

DIRECT TRANSFERRING RHIL-3 cDNA INTO MOUSE MUSCLE AND EXPRESSION IN VIVO

将含有人白细胞介素-3基因(hIL-3)cD-NA的真核表达质粒直接注射小鼠骨骼肌,观察hIL-3基因在小鼠肌肉及体内的分泌表达情况。原位分子杂交及免疫组化结果显示注射重组质粒后,第14大重组质粒全部进入到骨骼肌细胞内并有IL-3的表达。ELISA结果显示,注射重组质粒DNA后,第21天小鼠血液中IL-3的含量最高,约有67ng/ml,第14天次之,约51ng/ml,第28天和第7天相近,约34ng/ml。生物学活性检测结果表明,实验组小鼠血清有维持IL-3依赖的TF-1细胞株生长的作用。表明重组质粒DNA直接注射骨骼肌后进入到肌细胞内并表达IL-3,进入血液,而且表达产物有生物学活性。

The naked cucaryotic expression plasmid pKG5-IL-3 which contains hIL-3 cDNA, was di-rectly injected into the BALB/C mice's quadriceps muscle The results of in situ hybridization and immunohistro-chemistry showed that pKG5-IL-3 entered the muscle celis and expressed IL-3. The assay of ELISA showed that expressed hIL-3 was sccreted into blood The highest Ievel of IL-3 in serum was about 67 ng/ml on 21 st day after pKC5-IL-3 injection Bioactivity assay showed that the serum of experimental groups of mice was able to support the growth of the hIL-3 dependent TF-1 cells. These results suggest that the pKG5-IL-3 injected into mice's ouadriceps muscle could enter mice's muscle celis and expressed active hIL-3.