摘要
目的:克隆牦牛β-乳球蛋白(BLG)基因5’调控成分(3.5kb)。方法:利用PCR方法克隆了牦牛BLG基因5’侧翼区,并进行生物信息学分析表明。结论:获得了3.5kb的BLG基因5’调控成分,其中包括5’侧翼区(2 472bp),第一外显子及第一内含子(1 066bp)。该序列与牛、绵羊和山羊的同源性分别为98.42%、89.61%和84.41%;平均GC含量为49.3%;存在一个CpG岛;可能存在一个启动子位点,位于起始密码子前-83bp处;AliBaba2.1分析发现该区域有47种潜在的转录因子结合位点,其中包括乳蛋白结合因子(MPBF)、乳腺活化因子(MAF)、糖皮质激素受体(GR)、雌激素受体(ER)、核因子1(NF-1)等结合位点,提示该调控成分可用于构建乳腺特异性表达载体。结果:成功克隆出BLG基因5’侧翼区,为构建转基因牦牛乳腺生物反应器的特异性表达载体奠定基础。
Objective：Obtain the yak β-lactoglobulin gene 5＇flanking fragment.Method：With PCR amplification,the yak β-lactoglobulin gene 5＇flanking fragment was cloned.And had been analyzed with bioinformatics methods.Result：The 5＇flanking fragment consisted in part of the gene upstream region about 2 472bp,the first exon and intron about 1 066bp.The result showed that the homology with cattle,sheep and goat BLG gene was 98.42%,89.61% and 84.41%,respectively.The average content of GC was 49.3%.It had a CpG island and a possible promoter site at-83bp.It was analyzed by AliBaba2.1,and found 47 sorts of potential transcription factor binding sites such as milk protein binding factor（MPBF）,mammary activating factor（MAF）,glucocorticoid receptor（GR）,estrogen receptor（ER）,nuclear factor 1（NF-1）.It suggested that the cloned regulatory element could be used to construct mammary gland specific expressional vector.Conclusion：The BLG gene 5＇flanking fragment was obtained which provided a base for constructing the specific expressional vector of transgenic yak＇s mammary gland bioreactor.
出处
《生物技术》
CAS
CSCD
北大核心
2011年第1期7-10,共4页
Biotechnology
基金
四川省学术和技术带头人培养基金项目资助
