Overexpression Vector Construction and Tissue Expression Analysis of HMOX1 Gene in Hezuo Pigs

doi:10.11924/j.issn.1000-6850.casb2021-0808

Abstract

Abstract:

In order to construct an overexpression vector of HMOX1 gene in Hezuo pigs and analyze its tissue expression characteristics, the complete CDS region sequences of HMOX1 gene were obtained by PCR amplification, and then were inserted into pcDNA3.1 (+) blank vector to construct pcDNA3.1-HMOX1 overexpression vector. At the same time, the expression of HMOX1 gene in the heart, liver, spleen, lung, kidney, stomach, duodenum and longissimus dorsi muscle of Hezuo pigs were detected by real-time quantitative PCR. The results showed that the completed CDS region of 924 bp of HMOX1 gene was successfully amplified and identified by double enzyme digestion and sequencing, which was successfully linked to the pcDNA3.1(+) overexpression vector. Results of tissue expression analysis showed that HMOX1 gene expression was the highest in Hezuo pig spleen tissue, followed by liver, lung, kidney and heart, and the lowest was in duodenum. These results can provide reference for further research on the function of HMOX1 gene in the process of adaptation of Hezuo pigs to high altitude and hypoxia.

Key words: Hezuo pigs, HMOX1 gene, vector construction, tissue expression

CLC Number:

S828.8+9

LI Jianrong, WANG Wei, LUO Dingguo, MA Xiaoxia, XU Meiling, GUN Shuangbao, YANG Qiaoli. Overexpression Vector Construction and Tissue Expression Analysis of HMOX1 Gene in Hezuo Pigs[J]. Chinese Agricultural Science Bulletin, 2022, 38(29): 140-145.

Figures/Tables 5

References 26

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基因名	序列(5’-3’)	大小/bp
HMOX1-CDS	F：CTAGCTAGCGAGCACTCACAGCCCAACAG	924
HMOX1-CDS	R：CCGCTCGAGCTCTCCAGGCAGAGTTCACG	924

基因名	序列(5’-3’)	大小/bp
HMOX1-CDS	F：CTAGCTAGCGAGCACTCACAGCCCAACAG	924
HMOX1-CDS	R：CCGCTCGAGCTCTCCAGGCAGAGTTCACG	924

引物名称	引物序列(5’-3’)	退火温度	长度	区域
HMOX1	F:GACATGGCCTTCTGGTATGGG	60	141 bp	348~368
HMOX1	R: CATGTAGCGGGTGTAGGCGT	60	141 bp	488~469
β-actin	F: ATATTGCTGCGCTCGTGGT	60	148 bp	142~160
β-actin	R: TAGGAGTCCTTCTGGCCCAT	60	148 bp	289~270

引物名称	引物序列(5’-3’)	退火温度	长度	区域
HMOX1	F:GACATGGCCTTCTGGTATGGG	60	141 bp	348~368
HMOX1	R: CATGTAGCGGGTGTAGGCGT	60	141 bp	488~469
β-actin	F: ATATTGCTGCGCTCGTGGT	60	148 bp	142~160
β-actin	R: TAGGAGTCCTTCTGGCCCAT	60	148 bp	289~270