Molecular cloning and characterization of double-stranded cDNA coding for bovine chymosin

Moir, Donald T.; Mao, Jen-i; Schumm, James W.; Vovis, Gerald F.; Alford, B L; Taunton-Rigby, Alison

doi:10.1016/0378-1119(82)90197-4

Cited by 60 publications

(19 citation statements)

References 28 publications

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“…4). It seems likely that the inverted repeat is an artifact of the snapback type, generated during cDNA synthesis (17,18), particularly since a DR a clone of the same library also displays a similar inverted repeat (K. Gustafsson, personal communication). Moreover, the second, but not the first, ATG codon is flanked by nucleotides identical to those of the consensus sequence for initiation sites (19).…”

mentioning

confidence: 99%

cDNA clone for the human invariant gamma chain of class II histocompatibility antigens and its implications for the protein structure.

Claesson¹,

Larhammar²,

Rask³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

145

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The invariant y chain is transitorily associated with class H histocompatibility antigens during intracellular transport. We have isolated and sequenced a DNA clone corresponding to the human y chain. mRNA hybridizing to the cDNA clone translated into a 33,000-dalton chain that associated specifically with class H antigen a and f3 chains. The y chain consists of 216 amino acids. The two N-linked carbohydrates are attached to asparagines 114 and 120. A continuous stretch of hydrophobic and neutral amino acids occurs in positions 31-56 from the NH2 terminus. This region seems to constitute the transmembrane portion of the polypeptide chain. The positions of the carbohydrate moieties and the putative transmembrane segment indicate that the NH2 terminus of the ychain resides on the cytoplasmic side of the membrane. Cell-free translations in conjunction with radiochemical amino acid sequence analyses suggest that the y chain lacks an

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mentioning

confidence: 99%

cDNA clone for the human invariant gamma chain of class II histocompatibility antigens and its implications for the protein structure.

Claesson¹,

Larhammar²,

Rask³

et al. 1983

Proc. Natl. Acad. Sci. U.S.A.

145

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“…Marie and Harold, (1996) [24,25] reported that the fusion protein was expressed in Escherichia coli and SDS-PAGE and the Western blot analysis of chymosin-treated cell lysates showed a pH-dependent cleavage of the fusion proteins. Fusion proteins were also bioselectively immobilized onto biotinylated controlled-pore glass beads and treated with chymosin.…”

Section: Discussionmentioning

confidence: 99%

“…However, Moir [23][24][25], stated that a full-length cDNA copy of the mRNA encoding calf chymosin (also known as rennin), a proteolytic enzyme with commercial importance in the manufacturing of cheese, has been cloned in an f1 bacteriophage vector. The nucleotide sequence of the cDNA was determined, and the translation of that sequence into amino acids predicts that the zymogen prochymosin is actually synthesized in vivo as preprochymosin with a 16 amino acid signal peptide.…”

Section: Discussionmentioning

confidence: 99%

Cloning and In Vitro-Transcription of Chymosin Gene in E. coli

El-Sohaimy¹,

Elsayed²,

Hafez³

et al. 2010

TONUTRAJ

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Chymosin, commonly known as rennin, is the main milk-coagulating enzyme available in rennet. RNA was extracted from the abomasum of a suckling calf water buffalo and was subjected to RT-PCR using degenerate primers to amplify 850bp of the chymosin gene. The sequence was aligned with 19 different mammals' chymosin genes. The sequence revealed that there is a similarity to them ranging from 64% to 98%. The purified recombinant proteins were obtained from the transformed E. coli and yeast. The clotting activity of both of the resulting proteins was examined compared to the commercial peers. It was noticed that the concentration of the purified protein ranged from 15,000 to 40,000 MCU. Therefore, the activity of the obtained proteins was the same and it was 105% when compared to the commercial peer. Having examined the cytotoxicity of the purified proteins, the results revealed no toxicity. We can conclude that the obtained recombinant protein is more active and safe even when expressed in bacteria rather than yeast.

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“…Several aspartic protease genes from molds and bacteria have been cloned and expressed (15,22,33,44). Bovine chymosin has been cloned and successfully expressed in Escherichia coli (11,18,25,27), in Bacillus spp. (19,28), in yeasts (14,24,38,42), and in filamentous fungi (7,40,41).…”

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confidence: 99%

Cloning and Expression of clt Genes Encoding Milk-Clotting Proteases from Myxococcus xanthus 422

Poza

Prieto-Alcedo

Sieiro

et al. 2004

Appl Environ Microbiol

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The screening of a gene library of the milk-clotting strain Myxococcus xanthus 422 constructed in Escherichia coli allowed the description of eight positive clones containing 26 open reading frames. Only three of them (cltA, cltB, and cltC) encoded proteins that exhibited intracellular milk-clotting ability in E. coli, Saccharomyces cerevisiae, and Pichia pastoris expression systems.Currently, there are three main types of rennins used by the cheese-making industry: (i) rennins extracted from the abomasum of suckling ruminants (such as chymosin [EC 3.4.23.4]), (ii) rennins prepared from microbial broths, and (iii) recombinant rennins. In general, animal rennins are not sufficient to cover world demands, and this fact has prompted research into both microbial and recombinant rennins. Although a variety of bacteria, yeasts, and fungi have been isolated as natural producers of milk-coagulating enzymes (3,10,13,26,32,45), only two genera are used worldwide in cheese production: Mucor (12, 39) and Endothia (8, 15). Several aspartic protease genes from molds and bacteria have been cloned and expressed (15,22,33,44). Bovine chymosin has been cloned and successfully expressed in

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Molecular cloning and characterization of double-stranded cDNA coding for bovine chymosin

Cited by 60 publications

References 28 publications

cDNA clone for the human invariant gamma chain of class II histocompatibility antigens and its implications for the protein structure.

cDNA clone for the human invariant gamma chain of class II histocompatibility antigens and its implications for the protein structure.

Cloning and In Vitro-Transcription of Chymosin Gene in E. coli

Cloning and Expression of clt Genes Encoding Milk-Clotting Proteases from Myxococcus xanthus 422

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