The aim of the study is to construct cDNA libraries from the normal liver and regeneration liver of rat by SMART (switching mechanism at 5' end of RNA transcript) technique and analyze their quality. The total RNA was separated from the normal liver and regeneration liver of rat and the frist-strand cDNA was synthesized through reverse transcription by a modified oligo (dT) primer (contained sfi IB site) while the SMART oligonucleotide (contained sfi IA site) was utilized as a template so that the first-strand cDNA could be extended over the 5' end of mRNA. The double-strand cDNA was amplified by LD-PCR (long-distance PCR) with the above two primers and then digested by sfi I (IA & IB) restriction, enzyme. After cDNA size fractionation through Chroma Spin 400 column, the double-strand cDNA was ligated into the sfi I-digested lambda TripIEx2 vector and then the recombinant DNA was packagedin vitro. The unamplified rat normal liver cDNA library consists of 1.3×10(7) pfu/ml, and regeneration liver cDNA library consists of 1.6×10(7) pfu/ml in which the percentage of recombinant clones both are about 100%. Through testing, the high quality cDNA libraries containing full-length cDNA of rat normal liver and regeneration liver have been constructed. The titer of the amplified cDNA library is 4.5×10(10) pfu/ml and 3.6×10(10) pfu/ml. the average exogenous inserts of the recombinants both are about 1.5 kb. These results show that the normal liver and regeneration liver of rat cDNA libraries both have an excellent quality and lay solid foundation to study liver functions and the mechanism of liver regeneration.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3454212PMC
http://dx.doi.org/10.1007/BF02894281DOI Listing

Publication Analysis

Top Keywords

normal liver
24
liver regeneration
24
regeneration liver
24
cdna libraries
16
liver
14
cdna
13
rat normal
12
liver rat
12
cdna library
12
contained sfi
8

Similar Publications

Background: Inflammation is a critical protective response in the body, essential for combating infections and healing injuries. However, chronic inflammation can be harmful and significantly contribute to the development and progression of chronic diseases, with macrophage-mediated responses being central to these processes. This study presents "SBR-Pel," a new therapeutic blend of Shinbaro tab (SBR), a traditional herbal formula, and pelubiprofen (Pel), a non-steroidal anti-inflammatory drug, and investigated their combined anti-inflammatory effects to create a treatment that both improves efficacy and reduces side effects.

View Article and Find Full Text PDF

Autophagy-related long non-coding RNA MIR210HG plays a therapeutic role in hepatocellular carcinoma.

Discov Oncol

January 2025

Hebei Key Laboratory of Laboratory Animal Science, Hebei Medical University, No. 361 East Zhongshan Road, Changan District, Shijiazhuang, 050017, China.

Objective: This study aimed to investigate the role of the autophagy-related long noncoding RNA (lncRNA) MIR210HG in hepatocellular carcinoma and its potential as a therapeutic target.

Methods: LncRNA MIR210HG expression and its correlation with survival outcomes in hepatocellular carcinoma patients were analyzed using data from The Cancer Genome Atlas (TCGA). Kaplan-Meier and Cox regression analyses were conducted to assess survival correlations.

View Article and Find Full Text PDF

Gut microbiota may modify the association between dietary polyphenol intake and circulating levels of hippuric acid: results from a 1-year longitudinal study in China.

Am J Clin Nutr

January 2025

Department of Nutrition, Center for Big Data and Population Health of IHM, The Second Affiliated Hospital of Anhui Medical University, School of Public Health, Anhui Medical University, Hefei, China. Electronic address:

Background: Hippuric acid (HA), a host-microbe co-metabolite, normally derives from gut microbial catabolism of dietary polyphenols.

Objectives: We investigated the potential interplay between dietary polyphenols and gut microbiota on circulating HA levels, and examined the associations between serum concentrations of HA and cardiometabolic risk markers.

Methods: In a 1-year cohort of 754 community-dwelling adults, serum HA and its precursor [benzoic acid (BA)] and fecal microbiota were assayed using liquid chromatography-tandem mass spectrometry and 16S ribosomal RNA sequencing, respectively.

View Article and Find Full Text PDF

Discovery of thymoquinone analogues with high anthelmintic activity against monogenean infections in goldfish (Carassius auratus).

Vet Parasitol

January 2025

College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China; National Aquatic Animal Diseases Para-reference laboratory (HZAU), Wuhan 430070, China; Hubei Hongshan Laboratory, Wuhan 430070, China; Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention, Wuhan 430070, China. Electronic address:

Monogenean parasites are harmful pathogens in aquaculture systems. Current treatment strategies for monogenean infections are unsatisfactory, making the discovery of new drugs urgent. Thymoquinone (TQ), a natural monoterpene isolated from Nigella sativa L.

View Article and Find Full Text PDF

Comprehensive analysis of protein post-translational modifications reveals PTPN2-STAT1-AOX axis-mediated tumor progression in hepatocellular carcinomas.

Transl Oncol

January 2025

Zhejiang Provincial Key Laboratory of Pancreatic Disease, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China; Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China; Clinical Research Center of Hepatobiliary and Pancreatic Diseases, Zhejiang Province, China; Zhejiang University Cancer Center, Hangzhou, China. Electronic address:

Hepatocellular carcinoma (HCC) is a common malignant tumor. Although the proteomics of HCC is well studied, the landscape of post-translational modifications (PTMs) in HCC is poorly understood. The PTMs themselves and their crosstalk might be deeply involved in HCC development and progression.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!