异源MAR介导外源基因与家蚕受精卵核基质的体外结合

以携带人β干扰素基因5′上游MAR(Matrix Association regions)的质粒pCL为载体, 在其BamHI位点上正向插入天蚕(Antheraea ramamai)丝素基因核心区Af6.8而构建成质料pMAC。通过质粒pMAC、pAy6.8和pCL与家蚕(Bombyxmori)受精卵核基质的体外结合及其比较分析发现,在体外异源MA R能有效地介导外源基因与核基质结合。这为MAR作为真核基因转移、整合和表达的载体提供了一个初步的实验证据。 Abstract:We have constructed a plasmid pMAC by inserting the core region of Antheraen yamamai fibroin gene into the BamHI site of the plasmid pCL which contains the MAR upstream huIFN-βgene.In vitro association of pCL,pAy6.8 and pMAC with the nuclear matrix of Bombyx mori fertilized eggs indicated that pMAC and pCL could tightly bind to the nuclear matrix,but pAy 6.8 found much more ineffectively.This provides a preliminary evidence for MAR as a vector for eukaryotic genes tranfer,integration and expression.

In vitro Association of Exogenous Gene with the Nuclear Matrix of Bombyx mori Fertilised Eggs Mediated by Heterogeneous MAR

We have constructed a plasmid pMAC by inserting the core region of Antheraeayamamai fibroin gene into the BamHI site of the plasmid pCL which contains the MAR upstream huIFN-ggene. In vitro association of pCL, pAy6.8 and pMAC with the nuclear matrix of Bombyx mori fertilized eggs indicated that pMAC and pCL could tight1y bind to the nuclear mat fix, but pAy 6.8 found much more ineffectively. This provides a preliminary evidence for MAR as a vector for eukaryotic genes tranfer, integration and expression.