借助Red重组系统通過修復(GAp-repair)方式順利從BAC克隆RP42-254I13獲得包括αs1-、β-酪蛋白基因以及兩個基因之間的19.6kb序列在內的82kb的DNA序列(pBACLinksp-AD,在其它實驗中已已經成功獲得此重組質粒,見圖1),本試驗以這個82kb的重組質粒爲基礎,利用Red重組系統成功逐段敲除兩個牛酪蛋白基因之間的不同長度(19kb,14.5kb,10kb,5kb)的間隔序列以及αs1-酪蛋白基因的CDS區,獲得了缺失不同大小片斷的基因序列,以及缺失CDS區的αs1-酪蛋白基因,爲研究αs1-和β-酪蛋白巨大損失基因3^端側翼序列(19.6kb間隔序列在牛酪蛋白基因座控制區的定們奠定了技術基礎,並爲隨後插入外源基因,研究其表達功能,進行乳腺生物器的研究提供了方法。
Tumorigenesis is a complicated process which involves the activation of oncogenes and the repression of tumor suppressor etc. CHD5 belongs to CHD family and is a novel tumor suppressor identified by scientists in cold spring laboratory. Kinds of domains confer these CHDs more complexity and multifunctions. But there is only a little research about CHD5 by now, it has been found that CHD5 activates the ARF-P53 pathway via epigenetic modification to repress tumor and that CHD5 losted or mutant in most of neural system tumors. The more intensive research on this novel gene will be very useful for us to understand the mechanism of tumorigenesis and to develope tumor therapeutic agent. This article is to introduce the related fuction and signal pathway of the CHD family and CHD5 and make a prospect of CHD5 research.