%0 Journal Article
%T SLC22A12˲ʱתȾHEK293ϸ°ûÄ£Ð͵Ľ¨Á¢Óë¼ø¶¨<br>Establishment and identification of SLC22A12 transiently transfected HEK293 cell model
%A Áõ
%A ͤ
%A ÑîÓÑ»Ô
%A ÁõÏãÏã
%A ·ë
%A èª
%A ³Â
%A ÑÅ
%A Áõ´º»¨
%J Ö£ÖÝ´óѧѧ±¨£¨Ò½Ñ§°æ£©
%D 2018
%R 10.13705/j.issn.1671-6825.2017.12.109
%X Ä¿µÄ:¹¹½¨SLC22A12˲ʱתȾHEK293ϸ°ûÄ£ÐͲ¢½øÐмø¶¨¡£·½·¨:¹¹½¨GV142-SLC22A12ÖØ×éÖÊÁ£,²¢ÓÅ»¯ÖÊÁ£×ªÈ¾ÈËÅßÉöϸ°ûHEK293µÄÌõ¼þ¡£Í¨¹ýqRT-PCRºÍWestern blot¼ì²âSLC-HEK293ϸ°ûÖÐSLC22A12 mRNAºÍhURAT1µ°°×µÄ±í´ïÇé¿ö,½èÖú³¬¸ßЧҺÏàÉ«Æ×-´®ÁªÖÊÆ×·¨¿¼²ìÄòËáÄÆÉãÈ¡ÄÜÁ¦¡£½á¹û:Hind ¢óºÍEcoR ¢ñ˫øÇкͲâÐòÑéÖ¤ÏÔʾGV142-SLC22A12ÖÊÁ£¹¹½¨³É¹¦,×î¼ÑתȾÌõ¼þΪ2 ¦ÌgÖÊÁ£,ÊÔ¼Á(¦ÌL):ÖÊÁ£(¦Ìg)=3:1¡£ÓëHEK293ϸ°ûÏà±È,SLC-HEK293ϸ°ûÖÐSLC22A12 mRNAºÍhURAT1µ°°×±í´ïÁ¿Éϵ÷(P<0.001); ÄòËáÄÆÉãÈ¡ÄÜÁ¦¸üÇ¿(P<0.05)¡£½áÂÛ:³É¹¦½¨Á¢ÁËSLC22A12˲ʱתȾHEK293ϸ°ûÄ£ÐÍ¡£<br>Aim: To establish and identify a high SLC22A12 expression HEK293 cell line(SLC-HEK293).Methods: Recombinant plasmid GV142-SLC22A12 was constructed and the transfection condition was optimized.The expressions of SLC22A12 mRNA and hURAT1 protein in SLC-HEC293 cell were detected by qRT-PCR and Western blot, and the uptake of sodium urate was investigated by UPLC-MS/MS to confirm the construction of SLC-HEC293.Results: The plasmid of GV142-SLC22A12 was digested by Hind ¢ó and EcoR¢ñ and then was sequenced.The results showed that the GV142-SLC22A12 plasmid was constructed successfully.The optimal transfection conditions was 2 ¦Ìg plasmid with a 3:1 ratio of agent and plasmid. Compared with HEK293 cell transfected with GV142 plasmid, qRT-PCR and Western blot results showed that the expressions of SLC22A12 mRNA and hURAT1 protein in SLC-HEK293 cells were highly increased(P<0.001), and UPLC-MS/MS results showed that the uptake of sodium urate in SLC-HEK293 was also significantly higher(P<0.05).Conclusion: The transient transfection model of HEK293 cells highly expressing SLC22A12 has been established
%K SLC22A12
%K HEK293
%K תȾ&lt
%K br&gt
%K SLC22A12
%K HEK293
%K transfection
%U http://jms.zzu.edu.cn/oa/darticle.aspx?type=view&id=201805007