马铃薯甲虫热激蛋白基因<i>Ld-hsp70</i>的克隆及温度胁迫下的表达特性

马铃薯甲虫热激蛋白基因Ld-hsp70的克隆及温度胁迫下的表达特性

郑海霞,高玉林,张方梅,杨超霞,蒋健,朱勋,张云慧,李祥瑞

Cloning of Heat Shock Protein Gene Ld-hsp70 in Leptinotarsa decemlineata and Its Expression Characteristics under Temperature Stress

HaiXia ZHENG,YuLin GAO,FangMei ZHANG,ChaoXia YANG,Jian JIANG,Xun ZHU,YunHui ZHANG,XiangRui LI

表1 试验所用引物

Table1 Primers used in the study

引物名称 Primer name	引物序列 Primer sequence (5°-3°)	引物功能 Function of primers	退火温度 Annealing temperature (℃)
hsp70aF1	GCCATGAACCCCAATAACAC	hsp70a基因片段扩增 Amplification of hsp70a gene fragment	56.0
hsp70aR1	AGATGGTACCGGCATCTTTG		56.0
hsp70aF2	AGGAAGGACTTGACGAGCAA		56.0
hsp70aR2	TTTCTGCACCTTGGGGATAC		56.0
hsp70aF3	CAAGATCACCATCACCAACG		56.0
hsp70aR3	GATGGGGTTGCAGATGTTCT		56.0
hsp70aF4	CATACCTGGGCAAGACCG		56.0
hsp70aR4	CCAAAGACAGTGGCGTGA		56.0
hsp70bF1	GATGTTGGCACAGTCATTGG	hsp70b基因片段扩增 Amplification of hsp70b gene fragment	56.0
hsp70bR1	TGCTGAACAGTGGAGTCTGTG		56.0
hsp70bF2	AGCCAAGAGAGCCTTGTCTG		55.0
hsp70bR2	CAACGAGGACGATTTCATCA		55.0
hsp70cF1	GGCATTGATCTTGGAACCAC	hsp70c基因片段扩增 Amplification of hsp70c gene fragment	56.0
hsp70cR1	CTTCCTGGATCTTGGGATCA	hsp70c基因片段扩增 Amplification of hsp70c gene fragment	56.0
hsp70aF1	CAAGATCACCATCACCAACG	hsp70a 3° RACE扩增 3° RACE of hsp70a	56.0
hsp70bF1	TGATGAAATCGTCCTCGTT	hsp70b 3° RACE扩增 3° RACE of hsp70b	52.0
hsp70bF2	CAGCTCCCAGAGGTGTTC	hsp70b 3° RACE扩增 3° RACE of hsp70b	52.0
hsp70cF1	GCCAGGTAGCCATGAATC	hsp70c 3° RACE扩增 3° RACE of hsp70c	52.0
hsp70aFQ	ACGGTGACAAGTCTGAGGAGGT	实时荧光定量PCR qRT-PCR	60.0
hsp70aRQ	TGGTGGTGTTACGCTTGATGAG		60.0
hsp70bFQ	AGCAGCTATTGCCTATGGTTTG		60.0
hsp70bRQ	TTCCTGATGTCTTTGCCCTTC		60.0
hsp70cFQ	AACCAGTGGAGAAGGCTCTAACA		60.0
hsp70cRQ	TGTACTGCCGCACCATAAGC		60.0
GADPH-F	GATTCCACTCACGGACGATT	内参基因扩增 Amplification of the reference gene	60.0
GADPH-R	CATATTTGCCCCAAGGAATG		60.0
TUB-F	CTGCCTCTTTGAGGTTCGAC		60.0
TUB-R	TTACTGGGGCGTAAGTGACC		60.0