The genomic DNA Cosmid library was constructed from Nephila clavipes spider muscle using SuperCos 1 Cosmid as a vector.The title of library was >5×10 4 cfu/μg ligated DNA.On the basis of published sequence fr...The genomic DNA Cosmid library was constructed from Nephila clavipes spider muscle using SuperCos 1 Cosmid as a vector.The title of library was >5×10 4 cfu/μg ligated DNA.On the basis of published sequence from a partial cDNA sequence of the 3′end of the dragline silk gene, we designed and synthesized 3 oligonucleotides.Oligonucleotides were labeled with non radioactive digoxigenin dUTP and detected with chemilluminescent substrate.56 positive recombinants were screen from the Cosmid library using DIG Oligo 2 as a probe.DNA dot hybridization using DIG Oligo 1 and DIG Oligo 3 as the probes, respectively, 3 positive signals were identified from 56 colonies.They were appeared the same pattern when DNA from the colones digested by restriction enzymes.The spider dragline silk gene was confirmed again by Southern blot hybridization.展开更多
文摘The genomic DNA Cosmid library was constructed from Nephila clavipes spider muscle using SuperCos 1 Cosmid as a vector.The title of library was >5×10 4 cfu/μg ligated DNA.On the basis of published sequence from a partial cDNA sequence of the 3′end of the dragline silk gene, we designed and synthesized 3 oligonucleotides.Oligonucleotides were labeled with non radioactive digoxigenin dUTP and detected with chemilluminescent substrate.56 positive recombinants were screen from the Cosmid library using DIG Oligo 2 as a probe.DNA dot hybridization using DIG Oligo 1 and DIG Oligo 3 as the probes, respectively, 3 positive signals were identified from 56 colonies.They were appeared the same pattern when DNA from the colones digested by restriction enzymes.The spider dragline silk gene was confirmed again by Southern blot hybridization.
