Habenicht A et al. (1993), Molecular cloning of the large subunit of gluta...

XB-ART-22202

Biochim Biophys Acta 1993 Sep 23;11743:295-8.

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Molecular cloning of the large subunit of glutathione synthetase from Xenopus laevis embryos.

Habenicht A , Hille S , Knöchel W .

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We have isolated a cDNA clone from a Xenopus laevis tadpole cDNA library which probably codes for the large subunit of glutathione synthetase. The corresponding protein comprises 474 amino acids and shows a significant homology with the large subunit of glutathione synthetase of Schizosaccharomyces pombe. RNase protection experiments revealed that the gene is transcribed during oogenesis and that zygotic expression starts after midblastula transition. Transcripts are also detected in various adult tissues suggesting an ubiquitous distribution of the corresponding protein.

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Species referenced: Xenopus laevis
Genes referenced: gss pam sult1a1

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	Fig. 1. Nucleotide and predicted amino acid sequence of X. laet'is glutathione synthetase large subunit. The underlined region corresponds to the degeneraled oligonucleotide probe which was deduced from the Schizosaccharomyces pornbe amino acid sequence [5]. The 3' end shows a polyadenylation signal and a truncated poly(A) tail.
	Fig. 2. (a) Comparison of the X. laecis amino acid sequence (upper lane) with the amino acid sequence of the glutathione synthetase large subunit from Schizosa(charomyces pombe [5] (lower lane). Identical and PAM positions [6] are indicated. Deletions are marked by dashes. (b) Plot matrix of the Schizosaccharomyces pombe [5] gene sequence ( I 1725; Y-scale) with the Xem)pus cDNA sequence ( 1 - 150(J; X-scale). Window size: 30: allowed mismatches: 11.
	Fig. 3. RNase protection with RNAs from oocytes, embryos and adult tissues. Upper panel: a 333 bp Pstl/Dral fragment of X. laecis glutathione synthetase large subunit (GSLS) cDNA was subcloned into a pSPT 18. After Nhel cleavage within the vector and in vitro transcription, labelled antisense RNA was hybridized with different RNAs of indicated origins. RNase protected fragments were run on a denaturing polyacrylamide gel and submitted to autoradiography. Lower panel: RNase protection of the same RNAs with antisense IEF-Io~ RNA derived from a 311 bp Pcu II/ Pst I fragment of pXEF7 [7].