[1]杜玉杰,侯万儒.大熊猫RPL35A基因的克隆及超表达研究[J].西华师范大学学报(自然科学版),2018,39(04):377-382.[doi:10.16246/j.issn.1673-5072.2018.04.007]
 XU Hongxuan,LIU Xiaoqiang,CHANG Kai.Cloning and Overexpression of Ribosomal ProteinL35A Gene (RPL35A) from Giant Panda[J].Journal of China West Normal University(Natural Sciences),2018,39(04):377-382.[doi:10.16246/j.issn.1673-5072.2018.04.007]
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大熊猫RPL35A基因的克隆及超表达研究

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《西华师范大学学报(自然科学版)》[ISSN:1673-5072/CN:51-1699/N]

卷:
39
期数:
2018年04期
页码:
377-382
栏目:
出版日期:
2018-12-20

文章信息/Info

Title:

Cloning and Overexpression of Ribosomal ProteinL35A Gene (RPL35A) from Giant Panda

作者:

杜玉杰1侯万儒2

(1.湛江幼儿师范专科学校 科学教育系,广东 湛江  524048;2.西华师范大学 生命科学院,四川 南充  637009)

Author(s):

XU  Hongxuan  12  LIU  Xiaoqiang1CHANG  Kai  3

(1.SWUTAAHC Medicinal Plant Joint R&D Centre,School of Life Sciences,Southwest University,Chongqing 400715,China;2.Biology Teaching and Research Group,Chengdu Experiment Foreign Languages School,Chengdu Sichuan 611130,China;3.Department of Medical Laboratory,Chengdu Military General Hospital PLA,Chengdu Sichuan 610083,China)

关键词:

大熊猫RT- PCR降落PCRRPL35A克隆表达

Keywords:

giant pandaRT-PCRRPL35Acloningoverexpression

分类号:
Q786
DOI:
10.16246/j.issn.1673-5072.2018.04.007
文献标志码:
A
摘要:

探究大熊猫核糖体蛋白基因 RPL35A 及其编码蛋白的特点,以进一步研究其编码蛋白的功能,为深入开展大熊猫分子生物学研究及大熊猫基因保护等积累科学资料;本研究以大熊猫骨骼肌为材料,采用RTPCR和降落PCR方法克隆RPL35A基因的cDNA基因组序列,IPTG法诱导基因在BL21细胞系中的表达;大熊猫RPL35A基因的cDNA序列为337 bp,编码110个氨基酸;该基因的基因组序列长1 275 bp,包含2个外显子和1个内含子;RPL35A基因在BL21细胞系中的表达产物与组氨酸标签蛋白融合形成18 kDa的融合蛋白;该基因的cDNA序列、基因组序列中的外显子片段及其编码的蛋白序列与已报道的人、牛、小家鼠、褐家鼠等物种的对应序列同源性很高,蛋白质理化性质和功能位点基本相同,这说明该基因及其编码蛋白在进化过程中非常保守,可用于分子进化方面的研究,在物种进化关系研究中具有一定的参考意义和研究价值;同时,该基因可在原核生物中表达出具有活性的产物,可进一步开展纯化和蛋白质功能的研究。

Abstract:

To futher investigate the function of encoded protein,gather scientific data for molecular biology study of giant panda and protect gene resources of this endangered species,this paper explores the characteristics of ribosomal protein L35A gene (RPL35A) and its encoded protein.RT-PCR and touchdown PCR technology are used to clone the cDNA and the genomic sequence of RPL35A from giant panda and overexpressed in BL21 by IPTG.The results show that the length of cDNA is 333 bp encoding 110 amino acids and the genomic sequence is 1 275 bp in length possessing two exons and one introns.The expression protein of RPL35A in BL21 and N-terminally His-tagged protein give rise to the accumulation of an expected 18 kDa polypeptide fusion protein.The cDNA and its deduced amino acid sequence share a high homology with that of mammals reported,including Homo sapiens,Bos Taurus,Mus musculus and Rattus norvegicus.And their molecular weight as well as isoelectric point are basically the same.All of these illustrate that gene RPL35A and its encoded protein are very conservative in the process of evolution.These findings can be used in the study of molecular evolution.It is of certain reference significance and research value in phylogenic relationship.Meanwhile,its active overexpression in prokaryotic organism shows that further study on protein purification and its function can be conducted.

相似文献/References:

[1]李杨倩,唐博,周宏,等.历史皆伐林迹地中大熊猫的微生境选择——以栗子坪国家级自然保护区为例[J].西华师范大学学报(自然科学版),2018,39(04):345.[doi:10.16246/j.issn.1673-5072.2018.04.002]
 LI Yangqian,TANG Bo,ZHOU Hong,et al.Microhabitat Selection of Giant Pandas inHistorical Clear-cut Logging Forest—A Case Study of Liziping National Nature Reserve[J].Journal of China West Normal University(Natural Sciences),2018,39(04):345.[doi:10.16246/j.issn.1673-5072.2018.04.002]

备注/Memo

备注/Memo:

收稿日期:2018-01-23
基金项目:国家自然科学基金项目(31400016, 31200012)
作者简介:杜玉杰(1982—),女,河南周口人,讲师,主要从事生物化学和分子生物学研究。E-mail:xiaoshuidi156@126.com    
通信作者:侯万儒(1954—),男,四川阆中人,博士,教授,主要从事生物化学和分子生物学研究,E-mail:hwr@cwnu.edu.cn

更新日期/Last Update: 2018-12-20