[1]周 莹,王 楠,尹俊琦,等.大豆幼荚全长cDNA文库的构建及鉴定分析[J].大豆科学,2013,32(04):455-458.[doi:10.11861/j.issn.1000-9841.2013.04.0455]
　ZHOU Ying,WANG Nan,YIN Jun-qi,et al.Construction and Indentification of Fulllength cDNA Library from Young Pod of Soybean Jinong 18[J].Soybean Science,2013,32(04):455-458.[doi:10.11861/j.issn.1000-9841.2013.04.0455]
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大豆幼荚全长cDNA文库的构建及鉴定分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第32卷 期数: 2013年04期 页码: 455-458 栏目: 出版日期: 2013-08-25
Title:
Construction and Indentification of Fulllength cDNA Library from Young Pod of Soybean Jinong 18
作者:
?周 莹1; 王 楠1; 尹俊琦1; 姚 丹2; 马 建1; 曲 静1; 王丕武1
?1.吉林农业大学 农学院，吉林 长春 130118；2.吉林农业大学 生命科学学院，吉林 长春 130118
Author(s):
?ZHOU Ying1; WANG Nan1; YIN Jun-qi1; YAO Dan2; MA Jian1; QU Jing1; WANG Pi-wu1
?1.College of Agronomy,Jilin Agricultural University，Changchun 130118,China;2.College of Life Sciences,Jilin Agricultural University,Changchun 130118,China
关键词:
大豆; 幼荚; 全长cDNA文库; SMART技术
Keywords:
Soybean; Young pod; Fulllength cDNA library; SMART technology
分类号:
S336
DOI:
10.11861/j.issn.1000-9841.2013.04.0455
文献标志码:
A
摘要:
?????? 构建大豆幼荚全长cDNA文库，可为克隆与大豆产量相关基因，培育高产大豆新品种奠定基础。以吉农18大豆幼荚突变体为材料，提取大豆幼荚总RNA，反转录成单链cDNA，LDPCR方法合成双链cDNA；PCR产物经蛋白酶K消化、Sfil酶切后，采用CHROM SPIN400柱分级分离，回收500 bp以上的cDNA组分。以λTriplE×2载体连接并进行体外包装，构建吉农18大豆幼荚全长cDNA文库。经检测研究构建的大豆幼荚cDNA原始文库滴度达到1.5×106 pfu·mL-1，重组率达92％。扩增的文库滴定达到2.6×108 pfu·mL-1，重组cDNA片段长度在500 bp以上。结果显示构建的大豆幼荚全长cDNA文库符合高质量文库的标准，可用于进一步开展相关基因的克隆及分子生物学研究。
Abstract:
???? Construction of fulllength cDNA library from soybean young pod could lay foundation for cloning yield related genes.The total RNA from young pod of soybean Jinong 18 was separated and then transcribed into singlestrand cDNA,〖JP2〗which was synthesized into doublestrand cDNAs by LDPCR.Doublestrand cDNAs was digested by Proteinase K and Sfil,and fractionated by CHROMA SPIN400 columns.The ds cDNAs longer than 500 bp were collected and ligated to λTripIE×2,and then packaging reaction for recombinant bacteriophages was performed.The titer of the primary cDNA library was 1.5×106 pfu·mL-1,and the recombinant rate was 92％.The titer of amplified library was 2.6×108 pfu·mL-1and the insert size was more than 500 bp.Results showed that library titer and recombination rate met requirements of cDNA library,which would facilitate the functional genomics research of soybean.

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备注/Memo

?基金项目：吉林省科技厅科技支撑重点项目（20090203）；吉林省科技厅科技指导计划项目（201101111）。

更新日期/Last Update: 2014-02-14