Skip to main content
Log in

A family of serine protease genes expressed in adult buffalo fly (Haematobia irritans exigua)

  • Original Articles
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

Gene fragments encoding serine proteases expressed in adult buffalo fly (Haematobia irritans exigua) were amplified from cDNA using generic oligonucleotide PCR primers, based on conserved residues surrounding the active-site His and Ser amino acids found in all serine proteases. The PCR product consisted of a broad band extending from about 450 by to 520 bp, which suggested that the PCR product actually consisted of numerous DNA fragments of slightly variable sizes. Seventeen independent clones of these fragments, each with an insert of approximately 480 bp, were digested with HaeIII. Comparison of restriction fragment patterns indicated that 13 of these clones harboured different PCR products. This was confirmed by DNA sequence analysis of 9 clones. Each of the sequenced clones contained an open reading frame which included structurally conserved regions characteristic of the serine protease superfamily. This study reveals the expression of a large and highly variable repertoire of serine proteases in adult buffalo fly. Importantly, these data also demonstrate the utility of such an approach in obtaining DNA probes for use in further investigations of gene family organization and expression, as well as providing recombinant antigens in the form of fusion proteins which may be used as candidates for vaccine production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Allingham PG, Kerlin RL, Tellam RL, Briscoe SJ, Standfast HA (1992) Passage of host immunoglobulin across the mid-gut epithelium into the haemolymph of blood-fed buffalo flies (Haematobia irritans exigua). J Insect Physiol 38:9–17

    Google Scholar 

  • Barillas-Mury C, Graf R, Hagedorn HE, Wells MA (1991) cDNA and deduced amino acid sequences of a blood meal-induced trypsin from the mosquito Aedes aegypti. Insect Biochem 21:825–831

    Google Scholar 

  • Bean G, Seifert A, Macqueen A, Doube BM (1987) Effect of insecticide treatment for control of buffalo fly on weight gains of steers in coastal central Queensland. Aust J Exp Agric 27:329–334

    Google Scholar 

  • Ben-Yakir D (1989) Quantitative studies of host immunoglobulin in the haemolymph of ticks (Acari). J Med Entomol 26:243–246

    Google Scholar 

  • Borovsky D (1986) Proteolytic enzymes and blood digestion in the mosquito, Culex nigripalpus. Arch Insect Biochem Physiol 3:147–160

    Google Scholar 

  • Briegel H, Lea AO (1975) Relationship between protein and proteolytic activity in the midgut of mosquitoes. J Insect Physiol 21:1597–1604

    Google Scholar 

  • Brown JR, Hartley BS (1966) Location of disulphide bridges by diagonal paper electrophoresis. The disulphide bridges of bovine chymotrypsinogen A. Biochem J 101:214–228

    Google Scholar 

  • Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium-thyrocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159

    Article  CAS  PubMed  Google Scholar 

  • Davis CA, Riddell DC, Higgins MJ, Holden JJA, White BN (1985) A gene family in Drosophila melanogaster coding for trypsin-like enzymes. Nucleic Acids Research 13:6605–6619

    Google Scholar 

  • DeLotto R, Spierer P (1986) A gene required for the specification of dorsal-ventral pattern in Drosophila appears to encode a serine protease. Nature 323:688–692

    Google Scholar 

  • D'Onofrio G and Bernardi G (1992) A universal compositional correlation among codon positions. Gene 110:81–88

    Google Scholar 

  • Elizur A, Vacek AT, Howells AJ (1990) Cloning and characterisation of the white and topaz eye colour genes from the sheep blowfly Lucilia cuprina. J Mol Evol 30:347–358

    Google Scholar 

  • Emi M, Nakamura Y, Ogawa M, Yamamoto T, Nishide T, Utari T, Matsubara K (1986) Cloning, characterisation and nucleotide, sequences of his cDNAs encoding human pancreatic trypsinogens. Gene 41:305–310

    Google Scholar 

  • Ericson ML (1990) Quick DNA recovery from agarose gels by ultracentrifuge run. Trends Genet 6:278

    Google Scholar 

  • Felix CR, Betschart B, Billingsley PF, Freyvogel TA (1991) Postfeeding induction of trypsin in the midgut of Aedes aegypti L. (Diptera: Culicidae) is separable into two cellular phases. Insect Biochem 21:197–203

    Google Scholar 

  • Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791

    Google Scholar 

  • Fraser CM (1986) Merck Veterinary Manual 6th edn. Merck, Rahway, New Jersey

    Google Scholar 

  • Fujikawa K, Ghung DW, Hendrickson LE, Davie EW (1986) Amino acid sequence of human factor XI, a blood coagulation factor with four tandem repeats that are highly homologous with plasma prekallikrein. Biochemistry 25:2417–2424

    Google Scholar 

  • Gooding RE (1973) The digestive processes of haematophagous insects. IV. Secretion of trypsin by Aedes aegypti. Can I Entomol 105:599–603

    Google Scholar 

  • Graf R, Briegel H (1985) Isolation of trypsin isozymes from the mosquito Aedes aegypti. Insect Biochem 15:611–618

    Google Scholar 

  • Graf R, Briegel H (1989) The synthetic pathway of trypsin in the mosquito Aedes aegypti L. (Diptera: Culicidae) and in vitro stimulation in isolated midguts. Insect Biochem 19:129–137

    Google Scholar 

  • Graf R, Raikhel AS, Brown MR, Lea AO, Briegel H (1986) Mosquito trypsin: immunocytochemical localization in the midgut of blood-fed Aedes aegypti (L). Cell Tissue Res 245:19–27

    Google Scholar 

  • Grant GA, Henderson KO, Eisen AZ, Bradshaw RA (1980) Amino acid sequence of a collagenolytic protease from the hepatopancreas of the fiddler crab, Uca pugilator. Biochemistry 19:4653–4659

    Google Scholar 

  • Greer J (1990) Comparative modeling methods: Applications to the family of the mammalian serine proteases. Proteins: Structure, Function and Genetics 7:317–334

    Google Scholar 

  • Haber E, Quertermous T, Matsueda GR, Runge MS (1989) Innovative approaches to plasminogen activator therapy. Science 243:51–56

    Google Scholar 

  • Harris TJR (1987) Second generation plasminogen activators. Protein Engineering 1:449–458

    Google Scholar 

  • Higgins DG, Sharp PM (1989) Fast and sensitive multiple sequence alignments on a microcomputer. Comp Appl Biol Sci 5:151–153

    Google Scholar 

  • Higgins DL, Bennett WF (1990) Tissue plasminogen activator: the biochemistry and pharmacology of variants produced by mutagenesis. Annu Rev Pharmacol Toxicol 30:91–121

    Google Scholar 

  • Hogsette JA, Ruff JP (1986) Evaluation of flucythrinate and fenvalerate impregnated eartags and permethrin ear tapes for fly control on beef and dairy cattle in northwest Florida. J Econ Entomol 79:152–157

    Google Scholar 

  • Houseman JG, Campbell FC, Morrison PE (1987) A preliminary characterisation of digestive proteases in the posterior midgut of the stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae). Insect Biochem 17:213–218

    Google Scholar 

  • Inoue H, Nojima H, Okayama H (1990) High efficiency transformation of E. coli with plasmids. Gene 96:23–28

    Google Scholar 

  • Jany K-D, Haug H (1983) Amino acid sequence of the chymotryptic protease II from the larva of the hornet, Vespa crabro. FEBS Lett 158:98–102

    Google Scholar 

  • Jany K-D, Beckler G, Pleiderer G, Ishay J (1983) Amino acid sequence of an insect chymotrypsin from the larva of the hornet. Biochem Biophys Res Commun 110:1–7

    Google Scholar 

  • Jin Y, Anderson KV (1990) Dominant and recessive alleles of the Drosophila easter gene are point mutations at conserved sites in the serine protease catalytic domain. Cell 60:873–881

    Google Scholar 

  • Kemp DH, Agbede RIS, Johnston LAY, Gough JM (1986) Immunization of cattle against Boophilus microplus using extracts derived from adult female ticks: feeding and survival of the parasite on vaccinated cattle. Int J Parasitol 16:115–120

    Google Scholar 

  • Krätzschmar J, Haendler B, Langer G, Boidol W, Bringmann P, Alagon A, Donner P, Schleuning W-D (1991) The plasminogen activator family from the salivary gland of the vampire bat Desmodus rotundus. cloning and expression. Gene 105:229–237

    Google Scholar 

  • Kraut JU (1977) Serine proteases: structure and mechanism of catalysis. Annu Rev Biochem 46:331–358

    Google Scholar 

  • Lecroisey A, Gilles A-M, DeWolf A, Keil B (1987) Complete amino acid sequence of the collagenase from the insect Hypoderma lineatum. J Biol Chem 262:7546–7551

    Google Scholar 

  • Leytus SP, Loeb KR, Hagen FS, Kurachi K, Davie EW (1988) A novel trypsin-like serine protease hepsin with a putative transmembrane domain expressed by human liver and hepatoma cells. Biochemistry 27:1067–1074

    Google Scholar 

  • Moffatt MR, Lehane MJ (1990) Trypsin is stored as an inactive zymogen in the midgut of Stomoxys calcitrans. Insect Biochem 20:719–723

    Google Scholar 

  • Muta T, Hashimoto R, Miyata T, Nishimura H, Toh Y, Iwanaga S (1990) Proclotting enzyme from horseshoe crab haemocytes — cDNA cloning, disulphide locations and subcellular localisation. J Biol Chem 265:22426–22433

    Google Scholar 

  • Neurath H (1984) Evolution of proteolytic enzymes. Science 224:350–357

    Google Scholar 

  • Neurath H (1986) The versatility of proteolytic enzymes. J Cellular Biochem 32:35–49

    Google Scholar 

  • Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparisons. Proc Natl Acad Sci (USA) 85:2444–2448

    Google Scholar 

  • Ross MJ, Grossbard EB, Hotchkiss A, Higgins D, Anderson S (1988) Plasminogen activators. Ann Rep Med Chem 23:111–120

    Google Scholar 

  • Sakanari JA, Staunton CE, Eakin AE, Craik CS, McKerrow JH (1989) Serine proteases from nematode and protozoan parasites: Isolation of sequence homologues using generic molecular probes. Proc Natl Acad Sci (USA) 86:4863–4867

    Google Scholar 

  • Sambrook T, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. 2nd edn, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Seddon HR (1967) Diseases of domestic animals in Australia. 2. Arthropod infestations. Serv Publ No6 Dept Health Aust Smith DB, Johnson KS (1988) Single-step purification of polypeptides expressed in E. coli as fusions with glutathione S-transferase. Gene 67: 31-40

    Google Scholar 

  • Stevenson BT, Hagenbüchle, Wellaner PK (1986) sequence organisation and transcriptional regulation of the mouse elastase II and trypsin genes. Nucleic Acids Res 14:8307–8330

    Google Scholar 

  • Titani K, Sasagawa T, Woodbury RG, Ericsson LH, Dörsman H, Kraemer M, Neurath H, Zwilling R (1983) Amino acid sequence of crayfish (Astacus fluviatilis) trypsin If. Biochemistry 22:1459–1465

    Google Scholar 

  • Vaughan JA, Azad AF (1988) Passage of host immunoglobulin G from blood meal into haemolymph of selected mosquito species (Diptera: Culicidae). J Med Entomol 25:472–474

    Google Scholar 

  • Wikel SK (1988) Immunological control of haematophagous arthropod vectors: utilization of novel antigens. Vet Parasitol 29:235–264

    Google Scholar 

  • Willadsen P, Kemp DH (1988) Vaccination with ‘concealed antigens’: myth or reality? Parasitol Today 4:196–198

    Google Scholar 

  • Willadsen P, Riding G, McKenna R, Kemp DH, Tellam R, Nielsen JN, Lahnstein J, Cobon GS, Gough JM (1989) Immunological connrol of a parasitic arthropod: Identification of a protective antigen from Boophilus microplus. J Immunol 143:1346–1351

    Google Scholar 

  • Woodbury RG, Katunuma N, Kobayashi K, Titani K, Neurath H (1978) Covalent structure of a group-specific protease from rat small intestine. Biochemistry 17: 811–819

    Google Scholar 

  • Yun Y, Davis RL (1989) Levels of RNA from a family of putative serine protease genes are reduced in Drosophila melanogaster dunce mutants and are regulated by cyclic AMP. Mol Cell Biol 9:692–700

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by E.K.F. Bautz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Elvin, C.M., Whan, V. & Riddles, P.W. A family of serine protease genes expressed in adult buffalo fly (Haematobia irritans exigua). Molec. Gen. Genet. 240, 132–139 (1993). https://doi.org/10.1007/BF00276892

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00276892

Key words

Navigation