做分子实验,经常和不同的质粒打交道,了解各种质粒的图谱信息是必需的,今天就给大家介绍一下怎么查找质粒图谱。
方法一:使用 Vector NT 软件
Invitrogen 公司的这款软件绝对是分子生物学虫子们的福音。要想对质粒图谱了解更直观,安装这款软件是非常必要的。
这款软件的软件包里面会包括 Invitrogen 公司的所有质粒图谱信息和其他比较常见和经典的质粒图谱。
方法二:查找质粒图谱的网站
Vector Database(addgene)
地址:http://www.addgene.org/
这个网站很页面很人性化,以前叫做 lablife,现在网站做了整合,传送门 http://www.91bio.com/carriers-website-to-find/,比如查找 pRS 类质粒图谱,直接在搜索框输入 pRS,可以看到此类质粒一共有三十多个。
找到自己需要的质粒名称,点击进入,就可以看到质粒图谱了。
拿第一个质粒 pRSII413 举例,如上图,质粒图谱是不是很难看?对,我也觉得很难看,没关系,看见 view sequence 了吗?点击进入,我们就得到该质粒图谱的序列了。
Vector DB
网址:http://genome-www.stanford.edu/vectordb/vector.html
此网站页面比较简陋,但分类比较直观,总结和分类都比较完整。基本包括了所有表达系统的质粒信息。
不过唯一有一点不爽的就是,这个网站没有搜索栏,太不人性化了。那如何在那么多质粒信息中查找到我们所需要的质粒的信息呢?没关系,我们有网页字符查找功能,见红色标记部分。
方法是:网页工具栏编辑——–在此页上查找,然后输入你的质粒名称,就可以了。比如:我们输入 pRS,是不是就和搜索栏一样出现有用信息了?
点击我们需要的质粒进入下以页面,以质粒 pRS303 为例,如下图:有用信息除了对该质粒进行的一些描述性语言外,最重要的要算是红色标记部分了,Sequence Link 进入是该质粒的序列。NCBI Link 点击直接进入了 NCBI,如何在 NCBI 中得到质粒图谱,下面将进行详细解释。
NCBI
网址:http://www.ncbi.nlm.nih.gov/
重点介绍如何用 NCBI 查找质粒图谱信息。如下图,Search 下拉框中选中 Nucleotide,搜索栏输入质粒名称,拿 pRS303 举例:
Search 后,得到搜索结果,如下图,点击右边的 send to,选中 file、genebank 等选项,点击 Create File 选项,得到一个 gb 格式的文件,导入 vector 软件中,就得到了我们需要的质粒图谱了。
Google 及 Google 学术
Google 确实是个好东西,学生物的应该好好的学习下它的用法,虽然对于墙内有时总提示无法打开,但搜索的效率真的很高。
检索式子:质粒名 + map
质粒名 +sequence +filetype:txt or filetype:pdf
或者:进入 google,点击「图片搜索」,直接查找图谱。
其他的一些核酸数据库
欧洲分子生物学实验 EMBL:http://www.ebi.ac.uk/
方法三:一些重要试剂公司网页
其实也是一些网站,因为这些网站除了得到质粒图谱等信息,还可以得到关于质粒使用方面的信息,因此单独拿出来做一个分类。现在很多质粒,特别是应用比较广泛的质粒都一些公司商业化的质粒,基本是由一个质粒你就会联想到一个公司的名字。
比如简单的,克隆载体 pMD18-T,TaKaRa;pGM-T,天根;一些表达载体,使用最广泛的,pET 系列,Novagen 公司(默克)的;有双 MCS 的 Duet 系列载体,Novagen 的;毕赤酵母表达质粒 pPIC3.5k,pPIC9k,pPICZA,pPICZaA 等等,都是 Invitrogen 的;另外一些 pYES 酿酒酵母表达系统,也是 Invitrogen 的,因此知道常见的质粒是哪个公司的,去他们公司网站上肯定可以找到该质粒的相关信息。
在他们公司主页搜索栏直接搜索质粒名称,得到质粒序列,图谱什么的都不成问题,而且可以下到表达系统操作手册,原核表达看完 pET 表达系统操作手册,真核系统看完毕赤酵母表达系统。
方法四:如何查找经过改造过的质粒的质粒图谱
有些质粒是经过改造的,所以通过上述方法不能查询到相应信息。这时,可以在 google scholar 中输入质粒名称,可以直观地看哪些学者在何文章中使用了该质粒,从而可了解到质粒的来源或者籍此向作者咨询或索取。
google scholar: http://scholar.google.com/
另外介绍下如何通过文献查找经过改造的质粒的图谱信息,Kuhlman, T. E. and E. C. Cox (2010)。 「Site-specific chromosomal integration of large synthetic constructs.」 Nucleic Acids Res 38(6): e92.
这篇文献,介绍了一种大肠杆菌染色体整合的方法,文中介绍了三个质粒,是由作者自己改造了,如何查找到这三个质粒图谱呢?
首先在 PubMed 中搜索到该文献,如下图。
很多人找到文献,仅仅将文献下载下来就 OK 了,其实还有很多有用的信息,比如文章的 supplementary,还有如上图右边的标记部分,Nucleotide,protein 都是一些很重要的信息。点击右边的 Nucleotide,进入如下页面,文中涉及到的六个质粒序列全都有,点击进入,按照方法二中,NCBI 下载质粒图谱的方法就可以得到这几个质粒的图谱,当然,前提是该文章的作者已将载体信息上传 NCBI。
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