Limits...
Identification of a testis-enriched heat shock protein and fourteen members of Hsp70 family in the swamp eel.

He Y, Luo M, Yi M, Sheng Y, Cheng Y, Zhou R, Cheng H - PLoS ONE (2013)

Bottom Line: There are fourteen gene members in the Hsp70 family in the swamp eel genome.Hsp70 family, particularly Hspa8, has expanded in the species.One of the family members Hspa8b2 is predominantly expressed in testis of the swamp eel.

View Article: PubMed Central - PubMed

Affiliation: College of Fisheries, Huazhong Agricultural University, Wuhan, PR China.

ABSTRACT

Background: Gonad differentiation is one of the most important developmental events in vertebrates. Some heat shock proteins are associated with gonad development. Heat shock protein 70 (Hsp70) in the teleost fish and its roles in sex differentiation are poorly understood.

Methods and findings: We have identified a testis-enriched heat shock protein Hspa8b2 in the swamp eel using Western blot analysis and Mass Spectrometry (MS). Fourteen Hsp70 family genes were further identified in this species based on transcriptome information. The phylogenetic tree of Hsp70 family was constructed using the Maximum Likelihood method and their expression patterns in the swamp eel gonads were analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

Conclusion: There are fourteen gene members in the Hsp70 family in the swamp eel genome. Hsp70 family, particularly Hspa8, has expanded in the species. One of the family members Hspa8b2 is predominantly expressed in testis of the swamp eel.

Show MeSH

Related in: MedlinePlus

Western blot analysis of unknown testis-enriched protein in mouse and swamp eel.a. Western blot analysis using anti-NF-kB1 antibody in HeLa cells and the swamp eels showed two bands: 50kD (P50) and 105kD (P105) (stars denote the bands) in HeLa cells (two repeat samples), while only a dominant 70 KD band in testis of the swamp eel. β-actin protein was used as an internal control. Molecular weight sizes for proteins were shown on the right. b. Immunoprecipitation using anti-NF-kB1 antibody enriched the unknown testis-enriched protein in the swamp eel and mouse. Testis samples showed more obvious band of 70 kD in the swamp eel and mouse. Stars denote the band.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3672149&req=5

pone-0065269-g001: Western blot analysis of unknown testis-enriched protein in mouse and swamp eel.a. Western blot analysis using anti-NF-kB1 antibody in HeLa cells and the swamp eels showed two bands: 50kD (P50) and 105kD (P105) (stars denote the bands) in HeLa cells (two repeat samples), while only a dominant 70 KD band in testis of the swamp eel. β-actin protein was used as an internal control. Molecular weight sizes for proteins were shown on the right. b. Immunoprecipitation using anti-NF-kB1 antibody enriched the unknown testis-enriched protein in the swamp eel and mouse. Testis samples showed more obvious band of 70 kD in the swamp eel and mouse. Stars denote the band.

Mentions: Western blot analysis using anti-NF-kB1 antibody in HeLa cells showed normal two bands [50 kD (P50) and 105 kD (P105)], while only a testis-enriched band of 70 kD in the swamp eel was observed (Figure 1a). Further immunoprecipitation using anti-NF-kB1 antibody clearly showed the object unknown protein in testis samples of both the swamp eel and mouse (Figure 1b). The MALDI-TOF-TOF MS analysis was used to identify the proteins of both the swamp eel and mouse. The results indicated that the unknown testis-enriched protein was similar to Hsp70 family member Hspa2 or Hspa8 (Table 3), which had a conserved domain Hspa1–2_6–8-like_NBD (nucleotide-binding domain).


Identification of a testis-enriched heat shock protein and fourteen members of Hsp70 family in the swamp eel.

He Y, Luo M, Yi M, Sheng Y, Cheng Y, Zhou R, Cheng H - PLoS ONE (2013)

Western blot analysis of unknown testis-enriched protein in mouse and swamp eel.a. Western blot analysis using anti-NF-kB1 antibody in HeLa cells and the swamp eels showed two bands: 50kD (P50) and 105kD (P105) (stars denote the bands) in HeLa cells (two repeat samples), while only a dominant 70 KD band in testis of the swamp eel. β-actin protein was used as an internal control. Molecular weight sizes for proteins were shown on the right. b. Immunoprecipitation using anti-NF-kB1 antibody enriched the unknown testis-enriched protein in the swamp eel and mouse. Testis samples showed more obvious band of 70 kD in the swamp eel and mouse. Stars denote the band.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3672149&req=5

pone-0065269-g001: Western blot analysis of unknown testis-enriched protein in mouse and swamp eel.a. Western blot analysis using anti-NF-kB1 antibody in HeLa cells and the swamp eels showed two bands: 50kD (P50) and 105kD (P105) (stars denote the bands) in HeLa cells (two repeat samples), while only a dominant 70 KD band in testis of the swamp eel. β-actin protein was used as an internal control. Molecular weight sizes for proteins were shown on the right. b. Immunoprecipitation using anti-NF-kB1 antibody enriched the unknown testis-enriched protein in the swamp eel and mouse. Testis samples showed more obvious band of 70 kD in the swamp eel and mouse. Stars denote the band.
Mentions: Western blot analysis using anti-NF-kB1 antibody in HeLa cells showed normal two bands [50 kD (P50) and 105 kD (P105)], while only a testis-enriched band of 70 kD in the swamp eel was observed (Figure 1a). Further immunoprecipitation using anti-NF-kB1 antibody clearly showed the object unknown protein in testis samples of both the swamp eel and mouse (Figure 1b). The MALDI-TOF-TOF MS analysis was used to identify the proteins of both the swamp eel and mouse. The results indicated that the unknown testis-enriched protein was similar to Hsp70 family member Hspa2 or Hspa8 (Table 3), which had a conserved domain Hspa1–2_6–8-like_NBD (nucleotide-binding domain).

Bottom Line: There are fourteen gene members in the Hsp70 family in the swamp eel genome.Hsp70 family, particularly Hspa8, has expanded in the species.One of the family members Hspa8b2 is predominantly expressed in testis of the swamp eel.

View Article: PubMed Central - PubMed

Affiliation: College of Fisheries, Huazhong Agricultural University, Wuhan, PR China.

ABSTRACT

Background: Gonad differentiation is one of the most important developmental events in vertebrates. Some heat shock proteins are associated with gonad development. Heat shock protein 70 (Hsp70) in the teleost fish and its roles in sex differentiation are poorly understood.

Methods and findings: We have identified a testis-enriched heat shock protein Hspa8b2 in the swamp eel using Western blot analysis and Mass Spectrometry (MS). Fourteen Hsp70 family genes were further identified in this species based on transcriptome information. The phylogenetic tree of Hsp70 family was constructed using the Maximum Likelihood method and their expression patterns in the swamp eel gonads were analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

Conclusion: There are fourteen gene members in the Hsp70 family in the swamp eel genome. Hsp70 family, particularly Hspa8, has expanded in the species. One of the family members Hspa8b2 is predominantly expressed in testis of the swamp eel.

Show MeSH
Related in: MedlinePlus