Construction and identification of cDNA expression library of 3rd day larva of Echinococcus granulosus
10.16303/j.cnki.1005-4545.2025.02.18
- VernacularTitle:细粒棘球绦虫3 d幼虫cDNA表达文库的构建及鉴定
- Author:
Xiaoxia WU
1
;
Xiaolei LIU
;
Mingyuan LIU
;
Shumin SUN
;
Yongjun ZHAO
;
Jing DING
Author Information
1. 吉林大学动物医学学院/人畜共患传染病重症诊治全国重点实验室/人兽共患病研究教育部重点实验室,吉林长春 130062
- Publication Type:Journal Article
- Keywords:
Echinococcosis/hydatid disease;
Echinococcus granulosus;
3rd day larva;
cDNA expres-sion library
- From:
Chinese Journal of Veterinary Science
2025;45(2):305-311
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to construct and identify the cDNA expression library of 3rd day larva of Echinococcus granulosus.Echinococcus granulosus protoscolex was isolated and collected from the liver of a fresh infected sheep with cysts,and the experimental dogs were infected manually by mouth.On the 3rd day,approximately 1.5 meters of the duodenal jejunum was taken during the au-topsy,and the worm body was dislodged by swinging it in 38-40 ℃ physiological saline,and then the 3-day larvae of Echinococcus granulosus were collected.Total RNA was extracted by TRIzol reagent,mRNA was isolated and purified using PolyATtract? mRNA Isolation Systems.The cD-NA was synthesized by reverse transcription,ligated with pBluescript sk-vector after addition of a 5'junction and introduced into E.coli by electrotransformation to construct a cDNA expression li-brary for 3-day-old larvae of Echinococcus granulosus.The average length of insert cDNA frag-ments was 1.0-1.2 kb.The initial capacity of constructed library was 1.15×107 CFU/mL,titer of amplified library was 1× 1010 CFU/mL,recombination rate was 100％.The constructed cDNA ex-pression library and the inserted fragment size were suitable.This library is expected to be used for screening candidate genes and diagnostic antigen molecules of definitive hosts canine vaccines of Echinococcus granulosus.
