家蚕滞育激素基因的克隆

Cloning of the Diapause Hormone Gene of the Silkworm (Bombyx mori)

利用放射性同位素＾３２Ｐ－ｄＣＴＰ标记家蚕滞育激素ｃＤＮＡ作为探针，从家蚕基因库大约４×１０＾５个噬菌体斑中筛选得到３０个阳性克隆，其中３个克隆具有相同的限制性内切酶图谱，ｐｒｉｍｅｒ库列测定法，得到部分核苷酸序列，经比较，了这３个克隆确属滞育激素基因。基因组Ｓｕｒｔｈｅｒｎ分析，推定家蚕滞育激素基因属单基因编码。

A silkworm genomic library was screened with radiolabelled cDNA of Bombyx mori diapause hormone(Bom DH).Three positive clones were isolated from approximately 4×105 plaques,and subjected to further analyses.The DNA inserts excised from positive clones with SalⅠwere ligated into pBluescript KS(+)vector and grown in E.coli XL1 cells.The DNA fragments obtained by EcoRI digest form the insert were subcloned into the same vectors.The nucleotide sequence analysis showed that these clones involved the Bom DH gene.Genomic Southern analysis suggested that the Bom DH is encoded by a single gene. The present study which is the frist one on cloning of the Bom DH gene.