Li Zhao-lun, Tian Pu-xun, Xue Wu-jun
Renal Disease Center of First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 Nov;21(6):668-71.
To construct adenovirus expressing secretory human CD40L-Ig (sCD40 ligand Ig fusion protein).
The genes encoding the extracellular domain of human CD40L and IgG Fc were amplified with PCR and inserted into shuttle vector pAdTrack-CMV which then was transformed into E. coli pAdEasy-1-BJ5183 to produce recombinant Ad plasmid. The recombinant plasmid was digested with Pac I and transfected into 293 cells to generate recombinant adenovirus. The recombinant adenovirus was then used in mixed lymphocyte reaction (MLR) to test its function.
The recombinant adenovirus sCD40L-Ig was produced. It could inhibit the lymphocyte proliferation in MLR.
The sCD40L-Ig adenovirus is prepared successfully and its inhibition of MLR is confirmed.
构建表达分泌型人CD40L-Ig（sCD40配体Ig融合蛋白）的腺病毒。
采用PCR扩增编码人CD40L胞外区和IgG Fc的基因，并将其插入穿梭载体pAdTrack-CMV，然后将其转化至大肠杆菌pAdEasy-1-BJ5183中以产生重组腺病毒质粒。用Pac I酶切重组质粒并转染至293细胞中以产生重组腺病毒。然后将重组腺病毒用于混合淋巴细胞反应（MLR）以检测其功能。
产生了重组腺病毒sCD40L-Ig。它能够抑制MLR中的淋巴细胞增殖。
成功制备了sCD40L-Ig腺病毒，并证实了其对MLR的抑制作用。
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