Objective      To construct an expression plasmid vector carrying disrupted-in-schizophrenia 1 (DISC1) gene

and to determine the effect of DISC1 overexpression on membrane expansion of astrocytes.       Methods      Primers for the full-length of mouse DISC1 gene was designed and synthesized. The coding sequence of DISC1 gene was amplified by PCR from cDNA reversed from mouse white matter RNA

and then was inserted into a vector pEGFP-n1 using recombinant DNA technique. After verified by enzyme digestion and DNA sequencing

the recombinant plasmid named pEGFP-n1-DISC1 (DISC1 overexpression group) and the empty control pEGFP-n1 vector (control group) were transferred into astrocytes by electroporation

respectively. The expression of DISC1-GFP fusion protein in astrocytes was detected by Western blotting and immunofluorescent staining. The membrane expansion of astrocytes in the DISC1 overexpression group and control group were determined by analyzing the extended area of astrocytes with software Image-Pro Plus 5.0.       Results      The enzyme digestion and DNA sequencing confirmed that the coding sequence of mouse DISC1 was consistent with that in GenBank. The DISC1 was cloned into pEGFP-n1 vector in a right direction and named as pEGFP-n1-DISC1. The recombinant and control plasmids were effectively transferred into astrocytes with 40% of cells emitting green fluorescence after 60 h of transfection. The expression of DISC1 was only observed in cells transfected with pEGFP-n1-DISC1

and the membrane extended area of the DISC1 overexpression group significantly increased as compared with that of the control group (P0.05).       Conclusion      An exogenous expression plasmid vector carrying mouse DISC1 is successfully constructed

and DISC1 overexpression may induce the membrane expansion of astrocytes.