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Proteomic Analysis Of10-Hydroxy-2-Decanoic Acid Biosynthesis On Mandibular Gland Of Honeybee Worker(Apis Mellifera L.)

Posted on:2015-03-06Degree:MasterType:Thesis
Country:ChinaCandidate:X ZhangFull Text:PDF
GTID:2253330428973344Subject:Industry Technology and Engineering
Abstract/Summary:PDF Full Text Request
10-Hydroxy-2-Decanoic Acid (10-HDA) is one of the main active ingredient ofroyal jelly and a kind of trans fatty acids with various important physiologicalfunctions, such as antisepsis,immunoregulation and anti-tumor.10-HDA is mainlyproduced in mandibular gland of worker bees,but its biosynthesis mechanism is unclear.By analyzing10-HDA secretion regularity in woker mandibular gland,differentialproteins there at different day age was separated and identificated with the applicationof comparative protemics,screening releated proteins with10-HDA biosynthesis, toreveal its molecular mechanism for providing theoretical basis of increasing10-HDAproduction by using technology such as biology and genetic engineering.Workers(Apis mellifera ligustica) of shandong Changqing apiary was taken asexperimental animals in this paper, and10-HDA content in the head was detected byhigh performance liquid chromatography (HPLC) respectively on day0,5,10,15,20,25and30, researching secretion regularly of10-HDA with age. Gaschromatography-mass spectrome-try figure also confirmed that the sample extractedfrom worker head is indeed10-HDA solution. From day0to30,10-HDA secretionincrease gradually with the minimum at day0and the highest at day25, and it increasefastest from day20to25.Secretion regularity of10-HDA both in winter and summer is consistent and itscorresponding content in winter is more than it in summer. Found in the process ofexperimental test of each bee secretion by the content of10-HDA standard deviationis larger, so10-HDA content in different division of field bee head is also tested.theresult shows that the large deviation has nothing to do with the field of division oflabor.Experiments prove that worker bees mandibular gland is the source10-HDA,considering the experimental rigor and logical coherence, this paper also confirmedthis by contrast the10-HDA between the mandibular gland and worker bees headremoved the palate gland. Found10-HDA secretion the minmum at day0and thehighest at day25, firstly the worker bees mandibular gland corresponding morphologyat the two days of age was observed. mandibular gland at day0shape vacuole, and iseasy to break, not into a "V" shape, lustering.In constrast, at day25, it clearly shows"V" shape,and is lacklustre organizational entities, ivory or dark gray.scanning electron microscope the gland of25age is bigger than it at day0and full.Then29differencesprotein points of the mandibular gland were separated and identified bytwo-dimensional protein electrophoresis and mass spectrometry technology. most ofthe molecular weight concentrated in between20~70kd, and were mainly acidicprotein. carbohydrate and energy metabolism related protein was44.83%; exogenousprotein was13.79%;protein folding class accounted for10.34%;transcription andtranslation, cytoskeleton, oxidation resistance and the development of protein were6.90%; unknown protein accounted for3.45%.
Keywords/Search Tags:Apis mellifera ligustica, 10-HDA, Secretion regularity, Two-dimensionalelectr-ophoresis, Mass spectra
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