STAGE-SPECIFIC PROMOTER AND ITS CORE ACTIVE REGION OF BOMBYX MORI FOR REGULATION OF EARLY EMBRYONIC STAGE GENE EXPRESSION

摘要

PURPOSE: A promoter of silkworm embryonic stage-specific expression gene and an active region of the gene are provided to early determine transformation. CONSTITUTION: A silkworm embryonic stage-specific expression promoter is isolated from a silkworm and is denoted by sequence number 1. The promoter is denoted by sequence number 2 and is formed by deleting 268bp at sequence number 1. A maximum promoter active region of a silkworm embryonic stage-specific expression gene contains the promoter. A method for preparing the promoter comprises: a step of screening early embryo gene(EEG) through isolation of genetic resources with respect to time using Bombyx mori eggs and acquiring cDNA clone; a step of analyzing the characteristic and structure of total base sequence and predicted promoter region with respect to the clone of EEG-704 cDNA gene; a step of preparing deletion mutant constructs of 11 kinds of genes with respect to the promoter of EEG704 gene; a step of transducing each deletion mutant construct to pGL3-Basic carrier which do not has the promoter containing luciferase; a step of detecting luciferase activity with respect 11 kinds of deletion mutant constructs; and a step of comparing and analyzing luciferase activity with respect to p704A(1.4kb) and actin which have the strongest luciferase activity.