2010全国生物材料大会论文集之 (101).pdf

文档介绍：Cloning and Sequence Analysis of a cDNA of
P450 17α-hydroxylase/17,20-lyase(CYP17)
from Strongylocentrotus Nudus

Xiaoli Guo, Yang Liu, Xuemei Qiu, Xiuli Wang*
College of Life Sciences and Biotechnology,Dalian Fisheries University, Dalian 116023, China
*Corresponding author. E-mail: xiuliwang417@


Abstract—CYP17 (17α-hydroxylase/17, 20-lyase) has been II. MATERIALS AND METHODS
viewed as a critical enzyme for the biosynthesis of gonadal
A. Materials
and adrenal steroids in vertebrates. According to a reported
CYP17 gene of Strongylocentrotus purpuratus gene from The mature wild individuals of sea urchin (S. Nudus)
NCBI, three pairs of primers were designed to clone CYP17 was purchased from Heishijiao Market (Dalian, China)
gene of Strongylocentrotus nudus,using RT-PCR. Three and then dissected. The gonads were obtained and kept at
℃
fragments were assembled into on contig of 1569bp, -80 for RNA extraction.
including full CDS of 1482bp encoding 493 amino acid B. Isolation of RNA and sythesis of the first strand of
residues. Sequence analysis indicated the two sea urchin cDNA
CYP17 genes were mostly similar(97%) . But their amino Total RNA was extracted using Trizol reagent
acid identities were pared with vertebrates. Cloning (Tiangen, Beijing, China) from the gonads of sea urchin
of this CYP17 gene would be necessary to character