scholarly journals cDNA genes formed after infection with retroviral vector particles lack the hallmarks of natural processed pseudogenes.

1990 ◽  
Vol 10 (1) ◽  
pp. 68-74 ◽  
Author(s):  
R Dornburg ◽  
H M Temin
Keyword(s):  
Polymerase Chain Reaction ◽  
Full Length ◽  
Target Cells ◽  
Hygromycin B ◽  
Direct Repeats ◽  
Chromosomal Dna ◽  
Chain Reaction ◽  
Cis Acting ◽  
Processed Pseudogenes ◽  
Polymerase Chain

Retroviral proteins can encapsidate RNAs without retroviral cis-acting sequences. Such RNAs are reverse transcribed and inserted into the genomes of infected target cells to form cDNA genes. Previous investigations by Southern blot analysis of such cDNA genes suggested that they were truncated at the 3' and the 5' ends (R. Dornburg and H. M. Temin, Mol. Cell. Biol. 8:2328-2334, 1988). To analyze such cDNA genes further, we cloned three cDNA genes (derived from a hygromycin B phosphotransferase gene) in lambda vectors and analyzed them by DNA sequencing. We found that they did not correspond to the full-length mRNA: they were truncated at both the 3' and the 5' ends, did not contain a poly(A) tract, and were not flanked by direct repeats. The 3'-end junctions to chromosomal DNA of five more cDNA genes were amplified by polymerase chain reaction, cloned in pUC vectors, and sequenced. All of these cDNA genes had 3'-end truncations, and no poly(A) tracts were found. Further polymerase chain reaction experiments were performed to detect hygromycin B phosphotransferase cDNA genes with a poly(A) tract in DNA extracted from a pool of about 500 colonies of cells containing cDNA genes. No hygromycin B phosphotransferase cDNA gene with a poly(A) tract was found. Investigation of two preintegration sites by Southern analysis revealed that deletions were present in chromosomal DNA at the site of the integration of the cDNA genes. Naturally occurring processed pseudogenes correspond to the full-length mRNA, contain a poly(A) sequence, and are flanked by direct repeats. Our data indicate that cDNA genes formed by infection with retrovirus particles lack the hallmarks or natural processed pseudogenes. Thus, it appears that natural processed pseudogenes were not generated by retrovirus proteins.

cDNA genes formed after infection with retroviral vector particles lack the hallmarks of natural processed pseudogenes

1990 ◽  
Vol 10 (1) ◽  
pp. 68-74 ◽  
Author(s):  
R Dornburg ◽  
H M Temin
Keyword(s):  
Polymerase Chain Reaction ◽  
Full Length ◽  
Target Cells ◽  
Hygromycin B ◽  
Direct Repeats ◽  
Chromosomal Dna ◽  
Chain Reaction ◽  
Cis Acting ◽  
Processed Pseudogenes ◽  
Polymerase Chain

Retroviral proteins can encapsidate RNAs without retroviral cis-acting sequences. Such RNAs are reverse transcribed and inserted into the genomes of infected target cells to form cDNA genes. Previous investigations by Southern blot analysis of such cDNA genes suggested that they were truncated at the 3' and the 5' ends (R. Dornburg and H. M. Temin, Mol. Cell. Biol. 8:2328-2334, 1988). To analyze such cDNA genes further, we cloned three cDNA genes (derived from a hygromycin B phosphotransferase gene) in lambda vectors and analyzed them by DNA sequencing. We found that they did not correspond to the full-length mRNA: they were truncated at both the 3' and the 5' ends, did not contain a poly(A) tract, and were not flanked by direct repeats. The 3'-end junctions to chromosomal DNA of five more cDNA genes were amplified by polymerase chain reaction, cloned in pUC vectors, and sequenced. All of these cDNA genes had 3'-end truncations, and no poly(A) tracts were found. Further polymerase chain reaction experiments were performed to detect hygromycin B phosphotransferase cDNA genes with a poly(A) tract in DNA extracted from a pool of about 500 colonies of cells containing cDNA genes. No hygromycin B phosphotransferase cDNA gene with a poly(A) tract was found. Investigation of two preintegration sites by Southern analysis revealed that deletions were present in chromosomal DNA at the site of the integration of the cDNA genes. Naturally occurring processed pseudogenes correspond to the full-length mRNA, contain a poly(A) sequence, and are flanked by direct repeats. Our data indicate that cDNA genes formed by infection with retrovirus particles lack the hallmarks or natural processed pseudogenes. Thus, it appears that natural processed pseudogenes were not generated by retrovirus proteins.

Construction of a Full-Length Murine Proα2(I) Collagen cDNA by the Polymerase Chain Reaction

1991 ◽  
Vol 97 (6) ◽  
pp. 980-984 ◽  
Author(s):  
Charlotte L Phillips ◽  
Laura W Lever ◽  
Sheldon R Pinnell ◽  
Leigh D Quarles ◽  
Richard J Wenstrup
Keyword(s):  
Polymerase Chain Reaction ◽  
Full Length ◽  
Chain Reaction ◽  
Polymerase Chain

Amplification of Salmonella Chromosomal DNA Using the Polymerase Chain Reaction

1994 ◽  
Vol 38 (1) ◽  
pp. 119 ◽  
Author(s):  
Anh V. Nguyen ◽  
Mazhar I. Khan ◽  
Zhiqiang Lu
Keyword(s):  
Polymerase Chain Reaction ◽  
Chromosomal Dna ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Full-Length Sequence of Mouse Acupuncture-Induced 1-L (Aig1l) Gene Including Its Transcriptional Start Site

2011 ◽  
Vol 2011 ◽  
pp. 1-7
Author(s):  
Mika Ohta ◽  
Aki Sugano ◽  
Shuji Goto ◽  
Surini Yusoff ◽  
Yushi Hirota ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Polymerase Chain Reaction ◽  
Tissue Distribution ◽  
Cdna Sequence ◽  
Transcriptional Start Site ◽  
Full Length ◽  
Start Site ◽  
Chain Reaction ◽  
Polymerase Chain

We have been investigating the molecular efficacy of electroacupuncture (EA), which is one type of acupuncture therapy. In our previous molecular biological study of acupuncture, we found an EA-induced gene, named acupuncture-induced 1-L (Aig1l), in mouse skeletal muscle. The aims of this study consisted of identification of the full-length cDNA sequence ofAig1lincluding the transcriptional start site, determination of the tissue distribution ofAig1land analysis of the effect of EA onAig1lgene expression. We determined the complete cDNA sequence including the transcriptional start site via cDNA cloning with the cap site hunting method. We then analyzed the tissue distribution ofAig1lby means of northern blot analysis and real-time quantitative polymerase chain reaction. We used the semiquantitative reverse transcriptase-polymerase chain reaction to examine the effect of EA onAig1lgene expression. Our results showed that the complete cDNA sequence ofAig1lwas 6073 bp long, and the putative protein consisted of 962 amino acids. All seven tissues that we analyzed expressed theAig1lgene. In skeletal muscle, EA induced expression of theAig1lgene, with high expression observed after 3 hours of EA. Our findings thus suggest that theAig1lgene may play a key role in the molecular mechanisms of EA efficacy.

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