%0 Journal Article
%T Construction of recombinant plasmid of chicken calbindin-D28k<br>鸡CaBP-D28k基因重组质粒构建
%A YU Lu
%A WANG Ya-qian
%A ZHANG Shi-yuan
%A LI Yan-wu
%A LI Zhi-xue
%A <br>俞路
%A 王雅倩
%A 章世元
%A 李燕舞
%A 李治学
%J 浙江大学学报(农业与生命科学版)
%D 2008
%I Zhejiang University Press
%X To construct the eukaryotic expression vector of pCEP4-CaBP-D28k,a pair of primers was designed according to chicken calbindin-D28k(CaBP-D28k) gene sequence published in GenBank.Chicken CaBP-D28k gene was amplified by RT-PCR from chicken intestine total RNA extracts and was cloned into eukaryotic expression vector pCEP4,and transferred into COS-1,The gene expression was detected by indirect immunofluorescent assay(IFA).The result shows that the cloned CaBP-D28k gene,encoding 261 amino acids,was 789 bp in length which was 99.9% identity but only one nucleotide changed from G to T at site 760 compared with published sequence from GenBank database.The homology of CaBP-D28k gene sequence between chicken and other species such as frog,house mouse,human was 79.7%,78.2%,78.6%,respectively;the target segment in recombinant plasmid was characterized and confirmed by restrictive endonuclease assay as well as sequence analysis.The expression of CaBP-D28k could be detected in COS-1.This research provides a novel means to study regulation mechanism of calcium metabolism in chickens through pCEP4-CaBP-D28k.
%K chicken
%K calbindin-D28k gene
%K gene cloning
%K eukaryotic expression vector
%K indirect immunofluorescent assay(IFA)<br>鸡
%K CaBP-D28k
%K 基因克隆
%K 表达载体
%K 间接免疫荧光
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=03F54A49DE00578AA0E5DDF5BC021AA7&cid=298920A27C9BAA22346FCA384240FAA4&jid=466C05D88593309B3EB5CF34361E6426&aid=E43A18212A43F51806A3A013DB44A55B&yid=67289AFF6305E306&vid=339D79302DF62549&iid=B31275AF3241DB2D&sid=C7B13290323C226E&eid=2C20277AC27E4821&journal_id=1008-9209&journal_name=浙江大学学报(农业与生命科学版)&referenced_num=0&reference_num=19