东北黑熊不同基因组 DNA 模板来源的比较研究
收稿日期:2012-02-25
基金项目:校企合作研究项目(延大科合字(2009 第34 号))
作者简介:高建伟(1982-),男,博士研究生,研究方向为动物遗传育种。E-mail: 200603072@ybu. edu. cn
*通讯作者:鲁承,教授,博士生导师,研究方向为动物遗传育种。E-mail: clu@ybu. edu. cn
高建伟
1
,鲁 承
1*
,崔正云
1
,崔成都
1
,贾文影
2
,李 刚
3
( 1. 延边大学农学院动物医学系,吉林 延吉 133002;2. 科菲特饲料(齐齐哈尔)有限公司,黑龙江 齐齐哈尔 161200;
3. 长春大政药业科技有限公司,长春 130102 )
摘 要:文章主要探讨使用黑熊毛发作为试验材料替代其他生物样品进行遗传多样性研究的可行性,筛选满
足试验需求的最佳使用量。以延边白头山制药有限公司养熊场的 4 头成年东北黑熊为试验材料,分别5 次拾取 2 头
黑熊铁笼内散落的带有毛囊的 50、100 根毛发,对麻醉的 2 头黑熊抽取全血 3 mL 以及皮下结缔组织 1 g,并分别采
用不同方法提取基因组 DNA。以微卫星位点 ABB12 和 UT35 为引物,设置 20、50、100 ng·μL
-1
3 种模板浓度进行
PCR 扩增。结果显示,采用 50 ng·μL
-1
的模板浓度均可获得较好的 PCR 产物,采用聚丙烯酰胺凝胶电泳对扩增基
因具有较好的分辨效果。毛发 DNA、血液 DNA 和组织 DNA 的 PCR 产物数量和质量并无明显差异。毛发数量与所
提取的 DNA 浓度有一定关系。利用 15 个毛发 DNA 的 PCR 产物进行聚丙烯酰胺凝胶电泳,均可观察到清晰的条
带。研究结果进一步证明毛发 DNA 可作为东北黑熊遗传多样性研究的试验材料。
关键词:不同基因组 DNA 模板;微卫星;PCR 扩增
中图分类号:Q756 文献标志码:A 文章编号:1005-9369(2012)09-0090-06
Study on different sources of
U.t.ussuricus
genomic DNA template/GAO
Jianwei
1
, LU Cheng
1
, CUI Zhengyun
1
, CUI Chengdu
1
, JIA Wenying
2
, LI Gang
3
(1. Veterinary Depart-
ment, Agricultural College of Yanbian University, Yanji Jilin 133002, China; 2. Cofeed Feedmill(Qiqi-
har)Co., Ltd, Qiqihar Heilongjiang 161200, China; 3. Changchun Dazheng pharmaceutical science
and technology Co., Ltd, Changchun 130102, China)
Abstract: The research aimed to discuss the feasibility of make the genegic diversity research by
using
U.t.ussuricus
hair wholly as tested materials. With four adult U.t.ussuricus of Yanbian Baitou
mountain pharmacectic company, co,ltd bear breeding factory as tested materials, 50 and 100 hairs
were picked up five times in two bear cages and 3 mL whole blood were taken out and 1g subcutaneous
connective tissue were cut to make DNA extraction in different methods. The blood and subcutaneous
connective tissue were obtained, when the two bears were narcotized. With microsatellite primers
ABB12 and UT35 as primers,3template concentration including 20, 50, 100 ng·μL
-1
were set up to make
PCR amplification. Using 50 ng·μL
-1
as template concentration could obtain better PCR products, which
had better resolving effects on the amplification genes in polyacrylamide gel electrophoresis. The
quantity and quality of PCR products among hair, blood and tissue DNA samples had no obvious
difference. The hair number had suitable correlation with the concentration of DNA. When the PCR
products of 15 whole-hair samples were used in the native polyacrylamide gel electrophoresis,clear
bands could be observed. The research results further proved that DNA from whole-hair samples could
be taken as the materials for
U. t. ussuricus
genetic diversity research.
Key words: different genomic DNA template; microsatellite; PCR amplification
Journal of Northeast Agricultural University
2012 年9 月 Sep. 2012
东 北 农 业 大 学 学 报
第43 卷 第9 期 43(9): 90~95