Molecular and cytogenetic characterization of human albumin transgenic goat fibroblasts as a source of nuclei in somatic cloning

Abstract

Production of biopharmaceuticals in the mammary gland of domestic animals constitutes an important field of research in contemporary biotechnology. The objective of our investigations was to obtain goat transgenic fibroblast cells transfected with the assistance of the human albumin gene (pWAPhAlb) by the lipofection method for the needs of somatic cloning and to determine their molecular and cytogenetic characteristics. Transfection was carried out on 24 lines of goat fibroblasts and 19 lines in which integration of the introduced DNA was confirmed on the molecular level (PCR) were obtained. The method effectiveness reached 80%. Karyotype analysis (trypsin digestion, Giemsa stain) failed to show any changes in the chromosome number and structure(2n = 60, XX).

Full text

Ann. Anim. Sci., Vol. 11, No. 1 (2011) 61–70
MOLECULAR AND CYTOGENETIC CHARACTERIZATION
OF HUMAN ALBUMIN TRANSGENIC GOAT FIBROBLASTS
AS A SOURCE OF NUCLEI IN SOMATIC CLONING* *
11,22,
223,
31,2
1
2

3

Abstract
Production of biopharmaceuticals in the mammary gland of domestic animals constitutes an im-
portant field of research in contemporary biotechnology. The objective of our investigations was to
obtain goat transgenic fibroblast cells transfected with the assistance of the human albumin gene
(pWAPhAlb) by the lipofection method for the needs of somatic cloning and to determine their
molecular and cytogenetic characteristics. Transfection was carried out on 24 lines of goat fibrob-
lasts and 19 lines in which integration of the introduced DNA was confirmed on the molecular
level (PCR) were obtained. The method effectiveness reached 80%. Karyotype analysis (trypsin
digestion, Giemsa stain) failed to show any changes in the chromosome number and structure
(2n = 60, XX).
Key words: albumin, fibroblasts, transfection, nuclear transfer



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

             



A. Woźniak et al.
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






     

     
   



ALB

   


       


Material and methods

  


Characterization of human albumin transgenic goat fibroblasts 



  
2


32
   
           


  Aat 
Sph   
   
   
AatSph


  
    Sph  ClaI
    Sph  EcoR     

 Sph 
  ClaEcoR








  
    

           









A. Woźniak et al.
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






Characterization of human albumin transgenic goat fibroblasts 
 pWAPhAlb

       

      
pWAPhAlb




  




      

     

   






       
 
 


Results


         
          







A. Woźniak et al.
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
pWAPhAlb







pWAPhAlb


pWAPhAlb


Characterization of human albumin transgenic goat fibroblasts 
Discussion
      

            
     









           
     

 
    
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



  

    

   

  


A. Woźniak et al.
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
        
        


 

        


Competing interests

References

                                


  
           

                                             
                       


                                      
       


               

                                  


                               



             


                    

                                


                          


Characterization of human albumin transgenic goat fibroblasts 
           
                                

            


               

                                


            

                                          


                        

   

                  


                     


                          


         

                                      
   


                

                               
    
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

Molekularna i cytogenetyczna charakterystyka transgenicznych fibroblastów kozy transfeko-
wanych ludzkim genem albuminy jako źródła jąder komórkowych w klonowaniu somatycznym
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    



  

         

A. Woźniak et al.
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