IDENTIFICATION AND FUNCTIONAL ANALYSIS OF GENES RELATED TO LIVER METABOLISM IN SIBERIAN TIGER

Hongyi Yuan, Hairong Du, Ziao Yuan, Minghai Zjang*

College of wildlife and protected area, Northeast Forestry University, China, Harbin

Introduction: This study aimed to construct a cDNA expression library of the liver tissue of the Amur tiger to screen the related genes and provide the genetic basis for the animal model of medical research.

Method: In this study, the TRIzol method was employed to extract total RNA from frozen Amur tiger liver tissue, and a SMART cDNA library construction kit was used to construct the library. A total of 200 positive clones were randomly selected from the library under construction for sequencing analysis. The sequences were clustered and spliced using BLAST DNA STAR in GenBank to obtain the annotation of protein function in the database.

 Results: In the total RNA extracted at a concentration of 0.33 μg/μL (OD260/OD280 = 1.97), the size of the de novo constructed cDNA library was 2.68 × 106 PFU/mL, and the titer of the amplified library was 4.8 × 109 PFU/mL, with an average insert length of 0.78 kB and a recombination rate of 95.83%. Liver tissue is the largest digestive gland in organisms, representing the metabolic center of the body. The genetic information of the Siberian tiger was effectively preserved by constructing a cDNA library of the Siberian tiger liver. After successfully constructing the library, we screened out some important functional genes, such as SOD1, CTNNB1, LIF, and so forth, and carried out some analyses on them.

Conclusions: The cDNA library of the Amur tiger was successfully constructed, and the functional genes related to metabolism were screened out. Research on the protection of Siberian tiger genetic resources has very important scientific value and protection significance.