【摘要】：將多個(gè)可特異識(shí)別并結(jié)合DNA的鋅指單體串聯(lián)在一起即可構(gòu)成鋅指蛋白結(jié)構(gòu)域,若將其與不同的功能域因子融合,則會(huì)構(gòu)建出有很多用途的人工蛋白分子,從而實(shí)現(xiàn)對(duì)基因組的各種定點(diǎn)修飾或調(diào)控。以家蠶絲素重鏈基因?yàn)檠芯繉?duì)象,首先利用Zinc Finger Tools軟件篩選到了一個(gè)合適的靶標(biāo)位點(diǎn),再采用模塊組裝技術(shù),通過酶切、片段回收、連接、轉(zhuǎn)化等步驟成功將多個(gè)鋅指單體串聯(lián)在一起,構(gòu)建出一對(duì)靶向家蠶絲素重鏈基因特定位點(diǎn)的由4個(gè)鋅指單體組成的鋅指蛋白結(jié)構(gòu)域,這為將來定點(diǎn)修飾或改造家蠶絲素基因,進(jìn)一步提高蠶絲質(zhì)量和產(chǎn)量,以及大幅度提升家蠶生物反應(yīng)器對(duì)外源蛋白的表達(dá)能力打下了良好的基礎(chǔ)。
[Abstract]:Several zinc finger monomers that can specifically recognize and bind to DNA can be linked together to form zinc finger protein domains. If they are fused with different functional domain factors, artificial protein molecules with many uses will be constructed. In order to achieve a variety of genomes fixed-point modification or regulation. The silkworm fibroin heavy chain gene was used as the research object. A suitable target site was screened by using Zinc Finger Tools software, and then the module assembly technique was adopted, which was digested by enzyme, recovered and connected. Several zinc finger monomers were successfully linked together to construct a pair of zinc finger protein domains composed of 4 zinc finger monomers at specific sites of silkworm silk heavy chain gene. The further improvement of silk quality and yield, as well as the improvement of silkworm bioreactor to foreign protein expression ability laid a good foundation.
【作者單位】： 中山大學(xué)有害生物控制與資源利用國家重點(diǎn)實(shí)驗(yàn)室//昆蟲學(xué)研究所;
【基金】：廣東省科技計(jì)劃項(xiàng)目(2014A010107009)
【分類號(hào)】：Q78

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