海带配子体18S rRNA基因的克隆、序列分析及其作为内参基因的应用

doi:10.7668/hbnxb.2009.01.012

华北农学报 ›› 2009, Vol. 24 ›› Issue (1): 49-54. doi: 10.7668/hbnxb.2009.01.012

所属专题： 生物技术；

海带配子体18S rRNA基因的克隆、序列分析及其作为内参基因的应用

毕燕会, 邹丹燕, 周志刚

上海海洋大学水产与生命学院农业部水产种质资源与养殖生态重点开放实验室上海 200090

收稿日期:2008-09-08 出版日期:2009-01-28
通讯作者: 周志刚(1964)，安徽肥东人，教授，博士生导师，主要从事藻类生物学研究。
作者简介:毕燕会(1982-)女，山东菏泽人，博士，主要从事海洋生物技术研究
基金资助:
国家自然科学基金(30471328,30671627);上海市教育委员会重点研究项目(04KA02)

Cloning and Sequence Analysis of 18S rRNA Gene from Laminaria japonica Gametophytes and Its Application as an Internal Standard

BI Yan-hui, ZOU Dan-yan, ZHOU Zhi-gang

Key Laboratory of Aquatic Genetic Resources and Aquaculture Ecosystem The Ministry of Agriculture and College of Fisheries and Life Science Shanghai Ocean University Shanghai 200090 China

Received:2008-09-08 Published:2009-01-28

摘要/Abstract

摘要： 通过基因克隆和序列测定,获得了海带(Laminaria japonica)配子体的18S rRNA基因序列,其长度为1 716bp,GenBank登录号为EU293553。以蓝藻为外类群,采用邻接法(NJ)对分属9个门的藻类的18S rRNA基因序列进行系统发育分析。结果显示,褐藻门的海带与蓝藻门的念珠藻Nostoc sp.PCC7120、红藻门的日本凋毛藻(Griffithsia japoni-ca)、绿藻门的莱哈衣藻(Chlamydomonas reinhardtii)遗传距离依次为0.819,0.303和0.219 杂色藻类中,海带与隐藻门的蓝隐藻(Rhodomonas sp.CS24)遗传距离最大为0.201,与黄藻门的Heterosigma carterae遗传距离最近为0.103。提取不同光强条件下海带配子体RNA,利用18S rRNA基因作内参,通过半定量RT-PCR研究海带配子体光捕获蛋白基因lhcf6(Light-harvesting complex containing fucoxanthin)的不同光强条件下的表达量,揭示该基因随着光强增加而增加。

关键词: 海带, 18SrRNA基因, lhcf6, 半定量PCR

Abstract: 18S rRNfl gene sequence of Laminaria japonica was obtained by 18S rRNfl gene cloning and sequencing(CnBank accession number:EU293553).The phylogenetic analysis among 8 spieces andan outgro(Nostoc spPCC7120) were constructed based on 18S rRNfl gene sequences by neighborjoining(NT) method. Based ongenetic distance from L. japonica of Phaeophyta to Nostoc sp.PCC7120 of C}anophyta,Griffithsia japonicathe result,theof Rhocbphytaand Chlamydomonas reinhardtii of Chlorophyta,was 0. 819,0. 303 and 0. 219 in turn. While among chmmophyteslongest geneticshortest was 0.distance was 0. 201 from Laminaria japonica of Phaeophyta to Rhodomonas sp.CS24 of Cryptophyta,thethe103 from L. japonica to Heterosigma carterae of Xanthophyta. Using 18S rRNfl gene as internal standardsemi-quantitative RT-PCR result revealed the expression of lhcf6 gene increased with enhanced light intensity.

Key words: Laminaria.japonica, 18S rRNA gene, lhcf6, Semi-quantitative PCR

中图分类号:

S917.3

毕燕会, 邹丹燕, 周志刚. 海带配子体18S rRNA基因的克隆、序列分析及其作为内参基因的应用[J]. 华北农学报, 2009, 24(1): 49-54. doi: 10.7668/hbnxb.2009.01.012.

BI Yan-hui, ZOU Dan-yan, ZHOU Zhi-gang. Cloning and Sequence Analysis of 18S rRNA Gene from Laminaria japonica Gametophytes and Its Application as an Internal Standard[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(1): 49-54. doi: 10.7668/hbnxb.2009.01.012.

http://www.hbnxb.net/CN/Y2009/V24/I1/49

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[1]	谢玮怡, 毕燕会, 周志刚. 海带核基因组编码CbbX蛋白的原核表达与纯化[J]. 华北农学报, 2015, 30(2): 64-71.
[2]	宁璞, 毕燕会, 周志刚. *海带配子体光捕获蛋白基因lhcf6的启动子克隆与鉴定*[J]. 华北农学报, 2014, 29(1): 62-68.
[3]	毕燕会, 周志刚. *海带lhcf4基因克隆和配子体差异表达研究*[J]. 华北农学报, 2011, 26(6): 41-49.

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海带配子体18S rRNA基因的克隆、序列分析及其作为内参基因的应用

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