Cloning, sequencing and expression of cDNA encoding growth hormone from Indian catfish (Heteropneustes fossilis).
A tissue-specific cDNA library was constructed using polyA+ RNA from pituitary glands of the Indian catfish Heteropneustes fossilis (Bloch) and a cDNA clone encoding growth hormone (GH) was isolated. Using polymerase chain reaction (PCR) primers representing the conserved regions of fish GH sequences the 3' region of catfish GH cDNA (540 bp) was cloned by random amplification of cDNA ends and the clone was used as a probe to isolate recombinant phages carrying the full-length cDNA sequence. The full-length cDNA clone is 1132 bp in length, coding for an open reading frame (ORF) of 603 bp; the reading frame encodes a putative polypeptide of 200 amino acids including the signal sequence of 22 amino acids. The 5' and 3' untranslated regions of the cDNA are 58 bp and 456 bp long, respectively. The predicted amino acid sequence of H. fossils GH shared 98% homology with other catfishes. Mature GH protein was efficiently expressed in bacterial and zebrafish systems using appropriate expression vectors. The successful expression of the cloned GH cDNA of catfish confirms the functional viability of the clone.
Anathy
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Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai 625 021, India.
Venugopal
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Koteeswaran
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Pandian
T J
TJ
Mathavan
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S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002397:Catfishes / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000502:physiology; D015723:Gene Library; D049452:Green Fluorescent Proteins; D013006:Growth Hormone / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008164:Luminescent Proteins; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000737:chemistry / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016415:Sequence Alignment; D015027:Zebrafish / Q000196:embryology
D008164:Luminescent Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D013006:Growth Hormone