The Construction Of The Human HDAC1 Eukaryotic Expression Vector And The Modulation Of ERa And ERβ Protion Levels By HDAC1

Breast cancer is one of the most popular tumors of women. Estrogen play a critical role in normal breast development and function. The effects of estrogens are mediated by estrogen receptor (ERa and ERB), and ERs have been reported to bind various histones with high affinity. Acetylation is the most typical way of the activation of histones, which mediated by histone acetylase (HATs) and histone deacetylase(HDACs). In lacteal cell, ER control genetic expression by recruiting transcriptional cofactors, for HATs and HDACs example, which exhibit various enzymatic activities. ObjectiveConsequently, by constructing the pcDNA3-HA-hdacl, a HDAC1 eukaryotic expression vector, we were to study the interaction between HDAC1 and ERa and ERB in vitro and the effects of HDAC1 on the ERa and ERB at the protein level. So we can partly reveal the multiplication, differentiation and cancerization of lacteal gland cells. And we can make a grounding for looking for the target of anticancer drug preferably Methods(1)Extracting total RNA production from HepG2 cancer cell by TRIzol one-step method;(2)Synthetizing cDNA reversely transcription by using Promega reverse transcription kit, under the guide of operating instruction;(3)Amplifying the complete coding region array of HDAC1 gene by PCR;(4) Constructing the eukaryotic expression vector pcDNA3-HA-hdacl which...