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Expression of human transferrin can be regulated effectively by rabbit transferrin regulatory elements in transgenic mice

Expression of human transferrin can be regulated effectively...

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第八届全国医学生物化学与分子生物学第五届全国临床应用生物化学与分子生物学2013华东六省一市生物化学与分子生物学联合学术研讨会

作者： Jingbin Yan Xiuli Gong Shubiao Pan Xinbing Guo Zhaorui Ren Yitao Zeng Institute of Medical Genetics Children’s Hospital of Shanghai

human transferrin(hTF) belongs to iron-binding glycoprotein family which plays an important role in iron *** was also a significant factor for the culture of stem ***,the great demand of human transferrin could not be met because it was mainly isolated from the plasma ***,it was not expressed in milks of transgenic animal until now as a result of without proper mammary gland regulatory elements for the expression of hTF,although transgenic bioreactor was a good choice for the production of pharmaceutical ***,it would be useful to improving the expression level of hTF directed by efficient elements through analyzing the its regulatory *** this study,we prepared three expression construction named pcDNA3.1-2 EP-TFN,pcDNA3.1-EP-TFN and pcDNA3.1-RP-TFN,in which the expression of hTF gene was controlled by different rabbit transferring(rTF) regulatory cassette,and the differentiate expression of hTF was investigated in cell lines and transgenic ***,we also analyzed the methylation pattern of rabbit TF promoter in the various tissues of transgenic mice,which might regulate the specific expression of hTF gene in *** results indicated that hTF could be expressed effectively in the serum of transgenic mice when its expression was controlled by rTF promoter and enhancer,the highest expression level reached 913.6 g/***,rTF enhancer would repress the hTF expression in the milks when it was in a series of transgenic mice with rTF *** results also showed that tissue-specific expression of hTF gene was regulated by the methylation status of rTF promoter region,inverse relationship between methylation in rTF promoter and hTF expression level was significantly observed in the livers,hearts and muscles of transgenic *** change of methylation pattern on some transcription factor binding sites would be critical for the regulation of hTF expression in transgenic mice.

关键词： human transferrin Rabbit transferrin promoter Rabbit transferrin enhancer Methylation pattern Expression

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