Gene expression differences of regenerating rat liver in a short interval successive partial hepatectomy
4
Citation
62
Reference
10
Related Paper
Citation Trend
Abstract:
To identify the genes expressed differentially in the regenerating rat liver in a short interval successive partial hepatectomy (SISPH), and to analyze their expression profiles.Five hundred and fifty-one elements selected from subtractive cDNA libraries were conformed to a cDNA microarray (cDNA chip). An extensive gene expression analysis following 0-36-72-96-144 h SISPH was performed by microarray.Two hundred and sixteen elements were identified either up- or down-regulated more than 2-fold at one or more time points of SISPH. By cluster analysis and generalization analysis, 8 kinds of ramose gene expression clusters were generated in the SISPH. Of the 216 elements, 111 were up-regulated and 105 down-regulated. Except 99 unreported genes, 117 reported genes were categorized into 22 groups based on their biological functions. Comparison of the gene expression in SISPH with that after partial hepatectomy (PH) disclosed that 56 genes were specially altered in SISPH, and 160 genes were simultaneously up-regulated or down-regulated in SISPH and after PH, but in various amount and at different time points.Genes expressed consistently are far less than that intermittently; the genes strikingly increased are much less than that increased only 2-5 fold; the expression trends of most genes in SISPH and in PH are similar, but the expression of 56 genes is specifically altered in SISPH. Microarray combined with suppressive subtractive hybridization can in a large scale effectively identify the genes related to liver regeneration.Keywords:
Suppression subtractive hybridization
Liver Regeneration
Gene chip analysis
The cDNA from cerebellum of adult rat was taken as tester , and those from cerebellum of 1 day postnatal rat as driver . The suppression subtractive hybridization method was carried out. After cloning, a differential cDNA libraries was obtained. 54 clones were selected out and 56 ESTs were sequenced. 16 of 56 ESTs were identified to be novel ESTs and given the gene sequence numbers by GeneBank (BG946773-BG946788). Unknown ESTs can be selected after differentially expressed gene libraires was constructed with the method of suppression subtractive hybridization.
Suppression subtractive hybridization
Cloning (programming)
Cite
Citations (0)
To isolate peanut pod specially expressed genes,a cDNA library was constructed by using suppression subtractive hybridization(SSH).For subtractive hybridization,cDNA from the kernel of peanut was used as Driver,and cDNA from the pod of peanut was used as Tester.After two times of subtractive hybridization and two times of nested PCR,the products of last PCR amplification were inserted into pGEM-T Easy vectors and be transformed into the E.coli,a cDNA library of peanut pod was successfully constructed.89.2% of the colonies were white.The positive recombinants were confirmed by PCR method,83.5% of them showed single band.Those cDNA in the library can be used to isolate differentially expressed genes and promoters of peanut pod,also can be further studied for the purpose of understanding the molecular mechanism in the specially expression of pod genes.
Suppression subtractive hybridization
Cite
Citations (0)
Suppression subtractive hybridization
Representational difference analysis
Cite
Citations (118)
Suppression subtractive hybridization
Cucumis
Cite
Citations (4)
For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of Rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and fed male ticks were constructed using suppression subtractive hybridization (SSH). A total of 247 female expression sequence tags (ESTs) and 168 male ESTs were obtained from the two SSH cDNA libraries. It is predicted that 25 female ESTs and 44 female ESTs contain the 5′ and 3′ ends, respectively, and that 53 male ESTs and 74 male ESTs contain the 5′ and 3′ ends, respectively. To identify the subtraction rate of the two SSH cDNA libraries, the RT-PCR method was used to test 24 female ESTs and 21 male ESTs selected randomly but not repeatedly. The results showed that there were 13 upregulated or differentially expressed genes in the partially fed salivary gland of the female R. haemaphysaloides and that the differentially expressed rate was 54%. In addition, they indicated that there were 9 upregulated or differently expressed genes in the fed salivary gland of the male R. haemaphysaloides and that the differentially expressed rate was 43%. Putative translations of 141 (57%) female ESTs and 125 (74%) male ESTs had similarity to GenBank sequences, and 32 (23%) female ESTs and 29 (23%) male ESTs exhibited similarity to tick proteins, which showed that most of the proteins in the libraries were mainly related to the feeding blood physiology of the ticks.
Suppression subtractive hybridization
Cite
Citations (7)
The intestine of fish plays an important role in carbohydrate,protein,lipid metabolism and immunity.To obtain molecular biological information to the studies of some important physiological procession in grass carp.A subtractive cDNA library was constructed from the intestinal tract of grass carp experimentally infected with Aeromonas hydrophila by suppression subtractive hybridization(SSH).A total of 211 cDNA clones were partially sequenced and 147 ESTs obtained from the library.BLAST result showed that 147 of the intestinal ESTs could be ascribed to the transcriptional products of 97 identified genes,whereas 50 ESTs exhibited relatively low homology with unidentified sequences.Of the 97 known ESTs,there are 43 EST of cell defense-related,belonging to 13 genes.The successful constructed subtractive cDNA library will be essential for rapid isolation of immune-related genes of grass carp intestinal tissue and to further explore mucosal anti-infectious molecular mechanisms in fish.
Grass carp
Sequence (biology)
Cite
Citations (0)
Suppression subtractive hybridization
Verticillium dahliae
Gossypium barbadense
Verticillium wilt
Cite
Citations (40)
Objective To analyze differentially expressed gene in cerebellum of adult rat and to obtain a novel expressed sequence tag (EST).Methods The cDNA from cerebellum of adult rat was taken as tester, and those from cerebellum of 1 day postnatal rat as driver. The suppression subtractive hybridization method was carried out . After cloning, a differential cDNA libraries was obtained.Results 54 clones were selected out and 56 ESTs were sequenced. 16 of 56 ESTs were identified to be novel ESTs and given the gene sequence numbers by GeneBank (BG946773~BG946788).Conclusion Unknown ESTs can be selected after differentially expressed gene libraries was constructed with the method of suppression subtractive hybridization.
Suppression subtractive hybridization
Cloning (programming)
Identification
Cite
Citations (0)
Suppression subtractive hybridization
Northern blot
Cite
Citations (13)
OBJECTIVE: To expedite the subtractive screening of human hepatoma apoptotic cells cDNA library.METHODS: The method of subtractive hybridization combined with dot blot hybridization was adopted. First, minus cDNA probe was used to screen the cDNA library, the minus clones that did not hybridize with the minus cDNA probe were picked up as the source of the second grade dot blot hybridization. Secondly, two probes of the plus and minus cDNA were used to screen the clones;the clones that only hybridize with the plus cDNA probe were picked up as the source of the third grade dot blot hybridization. RESULTS: Four clones were obtained and the lengths of the inserted cDNA fragments were about 1.5 kb long. CONCLUSION: The results demonstrate this is a feasible, simple and quick method for subtractive screening of the cDNA library. (许昌泰)
Suppression subtractive hybridization
Dot blot
Southern blot
Northern blot
Cite
Citations (0)