[Amplification, cloning and sequence analysis of spider dragline silk cDNA].
Spider dragline silk is synthesized in special gland named major ampulate (MA) gland. The MA glands were dissected from the abdomen of the spiders Nephila clavata and the total RNA was extracted by the TRIZOL. The cDNA of dragline silk was amplificated by RT-PCR (reverse transcription polymerase chain reaction), multiplex PCR and cloned. PCR identification, restriction analysis and DNA sequence analysis were carried out to verify the recombinant plasmids. The codon usage frequencies of the cloned cDNA were added up, and the predicted amino acid sequence was compared with Spidroin2 of Nephila clavipes. Predicted secondary structure of the predicted amino-acid sequence was analysized by DNAStar software. All results showed that the cloned cDNA we got (GenBank Accession No. AF441245) was the very fragment of spider dragline silk Spidroin2 cDNA.
Zhang
Li-Shu
LS
Military Veterinary Institute, Quartermaster University of PLA, Changchun 130062, China.
Ma
He-Wen
HW
Lu
Yi-Ming
YM
Zhang
Yu-Jing
YJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D018076:DNA, Complementary / Q000737:chemistry*; D005349:Fibroins; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry / Q000235:genetics*; D013112:Spiders
D003062:Codon; D018076:DNA, Complementary; D011506:Proteins; C076815:spidroin 2; D005349:Fibroins